Benzimidazole derivatives: preparation and pharmaceutical applications

ABSTRACT

The present invention relates to hydroxamate compounds which are inhibitors of histone deacetylase. More particularly, the present invention relates to benzimidazole containing compounds and methods for their preparation. These compounds may be useful as medicaments for the treatment of proliferative disorders as well as other diseases involving, relating to or associated with dysregulation of histone deacetylase (HDAC).

FIELD OF THE INVENTION

The present invention relates to hydroxamate compounds that areinhibitors of histone deacetylase. More particularly, the presentinvention relates to benzimidazole containing compounds and methods fortheir preparation. These compounds may be useful as medicaments for thetreatment of proliferative disorders as well as other diseasesinvolving, relating to or associated with dysregulation of histonedeacetylase (HDAC).

BACKGROUND OF THE INVENTION

Local chromatin architecture is generally recognized as an importantfactor in the regulation of gene expression. The architecture ofchromatin, a protein-DNA complex, is strongly influenced bypost-translational modifications of the histones which are the proteincomponents. Reversible acetylation of histones is a key component in theregulation of gene expression by altering the accessibility oftranscription factors to DNA. In general, increased levels of histoneacetylation are associated with increased transcriptional activity,whereas decreased levels of acetylation are associated with repressionof gene expression [Wadem P. A. Hum. Mol. Genet. 10, 693-698 (2001), DeRuijter A. J. M. et al, Biochem. J., 370, 737-749 (2003)]. In normalcells, histone deacetylases (HDACs) and histone acetyltransferasetogether control the level of acetylation of histones to maintain abalance. Inhibition of HDACs results in the accumulation of acetylatedhistones, which results in a variety of cell type dependent cellularresponses, such as apoptosis, necrosis, differentiation, cell survival,inhibition of proliferation and cytostasis.

Inhibitors of HDAC have been studied for their therapeutic effects oncancer cells. For example, suberoylanilide hydroxamic acid (SAHA) is apotent inducer of differentiation and/or apoptosis in murineerythroleukemia, bladder, and myeloma cell lines [Richon V. M. et al,Proc. Natl. Acad. Sci. USA, 93: 5705-5708 (1996), Richon V. M. et al,Proc. Natl. Acad. Sci. USA, 95: 3003-3007 (1998)]. SAHA has been shownto suppress the growth of prostate cancer cells in vitro and in vivo[Butler L. M. et al, Cancer Res. 60, 5165-5170 (2000)]. Other inhibitorsof HDAC that have been widely studied for their anti-cancer activitiesare trichostatin A (TSA) and trapoxin B [Yoshida M. et al, J. Biol.Chem., 265, 17174 (1990), Kijima M. et al, J. Biol. Chem., 268, 22429(1993)]. Trichostatin A is a reversible inhibitor of mammalian HDAC.Trapoxin B is a cyclic tetrapeptide, which is an irreversible inhibitorof mammalian HDAC. However, due to the in vivo instability of thesecompounds they are less desirable as anti-cancer drugs. Recently, othersmall molecule HDAC inhibitors have become available for clinicalevaluation [U.S. Pat. No. 6,552,065]. Additional HDAC inhibitingcompounds have been reported in the literature [Bouchain G. et al, J.Med. Chem., 46, 820-830 (2003)] and patents [WO 03/066579A2, WO 01/38322A1]. The in vivo activity of such inhibitors can be directly monitoredby their ability to increase the amount of acetylated histones in thebiological sample. HDAC inhibitors have been reported to interfere withneurodegenerative processes, for instance, HDAC inhibitors arrestpolyglutamine-dependent neurodegeneration [Nature, 413(6857): 739-43, 18Oct., 2001]. In addition, HDAC inhibitors have also been known toinhibit production of cytokines such as TNF, IFN, IL-1 which are knownto be implicated in inflammatory diseases and/or immune systemdisorders. [J. Biol. Chem. 1990; 265(18): 10230-10237; Science, 1998;281: 1001-1005; Dinarello C. A. and Moldawer L. L. Proinflammatory andanti-inflammatory cytokines in rheumatoid arthritis. A primer forclinicians. 2^(nd) Edition, Amergen Inc., 2000].

Nevertheless, there is still a need to provide further HDAC inhibitorsthat would be expected to have useful, improved pharmaceuticalproperties such as anti-cancer agents.

SUMMARY OF THE INVENTION

In one aspect, the present invention provides compounds of the formula(I):

wherein:

R¹ is selected from the group consisting of: H, alkyl, alkenyl, alkynyl,haloalkyl, haloalkenyl, heteroalkyl, cycloalkyl, cycloalkenyl,heterocycloalkyl, heterocycloalkenyl, aryl, heteroaryl, cycloalkylalkyl,heterocycloalkylalkyl, arylalkyl, heteroarylalkyl, arylalkenyl,cycloalkylheteroalkyl, arylheteroalkyl, heterocycloalkylheteroalkyl,heteroarylheteroalkyl, hydroxy, hydroxyalkyl, alkoxy, alkoxyalkyl,alkoxyaryl, alkenyloxy, alkynyloxy, cycloalkylkoxy, heterocycloalkyloxy,aryloxy, heteroaryloxy, arylalkyloxy, amino, alkylamino, aminoalkyl,acylamino, arylamino, phenoxy, benzyloxy, COOH, alkoxycarbonyl,alkylaminocarbonyl, sulfonyl, alkylsulfonyl, alkylsulfinyl,arylsulfonyl, arylsulfinyl, aminosulfonyl, SR⁶ and acyl, each of whichmay be unsubstituted or substituted with one or more substituentsindependently selected from the group consisting of: halogen, ═O, ═S,—CN, —NO₂, —CF₃, —OCF₃, alkyl, alkenyl, alkynyl, haloalkyl, haloalkenyl,haloalkynyl, heteroalkyl, cycloalkyl, cycloalkenyl, heterocycloalkyl,heterocycloalkenyl, aryl, heteroaryl, hydroxy, hydroxyalkyl, alkoxy,alkoxyalkyl, alkoxyaryl, alkoxyheteroaryl, alkenyloxy, alkynyloxy,cycloalkyloxy, cycloalkenyloxy, heterocycloalkyloxy,heterocycloalkenyloxy, aryloxy, heteroaryloxy, arylalkyl,heteroarylalkyl, arylalkyloxy, amino, alkylamino, acylamino, aminoalkyl,arylamino, sulfonyl, alkylsulfonyl, arylsulfonyl, arylsulfinyl,aminosulfonyl, aminoalkyl, alkoxyalky, —COOH, —C(O)OR⁵, —COR⁵, —SH,—SR⁶, —OR⁶ and acyl;

or R¹=L;

R² is selected from the group consisting of: H, halogen, alkyl, alkenyl,alkynyl, haloalkyl, haloalkenyl, heteroalkyl, cycloalkyl, cycloalkenyl,heterocycloalkyl, heterocycloalkenyl, aryl, heteroaryl, cycloalkylalkyl,heterocycloalkylalkyl, arylalkyl, heteroarylalkyl, arylalkenyl,cycloalkylheteroalkyl, heterocycloalkylheteroalkyl,heteroarylheteroalkyl, arylheteroalkyl hydroxy, hydroxyalkyl, alkoxy,alkoxyalkyl, alkoxyaryl, alkenyloxy, alkynyloxy, cycloalkylkoxy,heterocycloalkyloxy, aryloxy, heteroaryloxy, arylalkyloxy, amino,alkylamino, aminoalkyl, acylamino, arylamino, phenoxy, benzyloxy, COOH,alkoxycarbonyl, alkylaminocarbonyl, sulfonyl, alkylsulfonyl,alkylsulfinyl, arylsulfonyl, arylsulfinyl, aminosulfonyl, SR⁶ and acyl,each of which may be unsubstituted or substituted with one or moresubstituents independently selected from the group consisting of:halogen, ═O, ═S, —CN, —NO₂, —CF₃, —OCF₃, alkyl, alkenyl, alkynyl,haloalkyl, haloalkenyl, haloalkynyl, heteroalkyl, cycloalkyl,cycloalkenyl, heterocycloalkyl, heterocycloalkenyl, aryl, heteroaryl,hydroxy, hydroxyalkyl, alkoxy, alkoxyalkyl, alkoxyaryl,alkoxyheteroaryl, alkenyloxy, alkynyloxy, cycloalkyloxy,cycloalkenyloxy, heterocycloalkyloxy, heterocycloalkenyloxy, aryloxy,heteroaryloxy, arylalkyl, heteroarylalkyl, arylalkyloxy, amino,alkylamino, acylamino, aminoalkyl, arylamino, sulfonyl, alkylsulfonyl,arylsulfonyl, aminosulfonyl, aminoalkyl, alkoxyalky, —COOH, —COR⁵,—C(O)OR⁵, —SH, —SR⁶, —OR⁶ and acyl;

or R²=L;

R³ is selected from the group consisting of H, C₁-C₆ alkyl, and acyl; ora metal ion selected from sodium, calcium, magnesium;

X and Y are the same or different and are independently selected fromthe group consisting of: H, halogen, —CN, —NO₂, —CF₃, —OCF₃, alkyl,alkenyl, alkynyl, haloalkyl, haloalkenyl, haloalkynyl, heteroalkyl,cycloalkyl, cycloalkenyl, heterocycloalkyl, heterocycloalkenyl, aryl,heteroaryl, hydroxy, hydroxyalkyl, alkoxy, alkoxyalkyl, alkoxyaryl,alkoxyheteroaryl, alkenyloxy, alkynyloxy, cycloalkyloxy,cycloalkenyloxy, heterocycloalkyloxy, heterocycloalkenyloxy, aryloxy,heteroaryloxy, arylalkyl, heteroarylalkyl, arylalkyloxy, amino,alkylamino, acylamino, aminoalkyl, arylamino, sulfonyl, alkylsulfonyl,arylsulfonyl, aminosulfonyl, aminoalkyl, alkoxyalky, —COOH, —C(O)OR⁵,—COR⁵, —SH, —SR⁶, —OR⁶, acyl and —NR⁷R⁸;

R⁴ is selected from the group consisting of: H, alkyl, alkenyl, alkynyl,haloalkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl,cycloalkylalkyl, heterocycloalkylalkyl, arylalkyl, heteroarylalkyl andacyl;

Each R⁵ is independently selected from the group consisting of: H,alkyl, alkenyl, alkynyl, haloalkyl, heteroalkyl, cycloalkyl,heterocycloalkyl, aryl, heteroaryl, cycloalkylalkyl,heterocycloalkylalkyl, arylalkyl, heteroarylalkyl and acyl;

Each R⁶ is independently selected from the group consisting of: H,alkyl, alkenyl, alkynyl, haloalkyl, heteroalkyl, cycloalkyl,heterocycloalkyl, aryl, heteroaryl, cycloalkylalkyl,heterocycloalkylalkyl, arylalkyl, heteroarylalkyl and acyl;

Each R⁷ and R⁸ are each independently selected from the group consistingof: H, alkyl, alkenyl, alkynyl, haloalkyl, heteroalkyl, cycloalkyl,heterocycloalkyl, aryl, heteroaryl, cycloalkylalkyl,heterocycloalkylalkyl, arylalkyl, heteroarylalkyl and acyl;

L is selected from the group consisting of:

-   -   a) L=Cy-L¹-W—    -   Wherein        -   Cy is C₁-C₁₅ alkyl, aminoalkyl, heterocycloalkyl,            cycloalkyl, aryl, aryloxy or heteroaryl, any of which may be            optionally substituted one or more substituents            independently selected from the group consisting of:            halogen, ═O, ═S, —CN, —NO₂, —CF₃, —OCF₃, alkyl, alkenyl,            alkynyl, haloalkyl, haloalkenyl, haloalkynyl, heteroalkyl,            cycloalkyl, cycloalkenyl, heterocycloalkyl,            heterocycloalkenyl, aryl, heteroaryl, hydroxy, hydroxyalkyl,            alkoxy, alkoxyalkyl, alkoxyaryl, alkoxyheteroaryl,            alkenyloxy, alkynyloxy, cycloalkyloxy, cycloalkenyloxy,            heterocycloalkyloxy, heterocycloalkenyloxy, aryloxy,            heteroaryloxy, arylalkyl, heteroarylalkyl, arylalkyloxy,            amino, alkylamino, acylamino, aminoalkyl, arylamino,            sulfonyl, alkylsulfonyl, arylsulfonyl, aminosulfonyl,            aminoalkyl, alkoxyalky, —COOH, —C(O)OR⁵, —COR⁵, —SH, —SR⁶,            —OR⁶ and acyl.        -   L¹ is selected from the group consisting of C₁-C₅ alkyl,            which may be optionally substituted with one or more            substituents independently selected from the group            consisting of: halogen; ═O; ═S; —CN; —NO₂; alkyl, alkoxy,            acylamino, and alkylamino;        -   W is selected from the group consisting of a single bond,            —O—, —S—, —S(O)—, —S(O)₂—, —N(R⁹)—, —C(O)N(R⁹)—, —SO₂N(R⁹)—,            N(R⁹)C(O)—, N(R⁹)SO₂—, and —N(R⁹)—C(O)—N(R¹⁰)—;    -   b) L=Cy-L¹-W-L²    -   Wherein,        -   Cy is C₁-C₁₅ alkyl, aminoalkyl, heterocycloalkyl,            cycloalkyl, aryl, aryloxy or heteroaryl, any of which may be            optionally substituted one or more substituents            independently selected from the group consisting of:            halogen, ═O, ═S, —CN, —NO₂, —CF₃, —OCF₃, alkyl, alkenyl,            alkynyl, haloalkyl, haloalkenyl, haloalkynyl, heteroalkyl,            cycloalkyl, cycloalkenyl, heterocycloalkyl,            heterocycloalkenyl, aryl, heteroaryl, hydroxy, hydroxyalkyl,            alkoxy, alkoxyalkyl, alkoxyaryl, alkoxyheteroaryl,            alkenyloxy, alkynyloxy, cycloalkyloxy, cycloalkenyloxy,            heterocycloalkyloxy, heterocycloalkenyloxy, aryloxy,            heteroaryloxy, arylalkyl, heteroarylalkyl, arylalkyloxy,            amino, alkylamino, acylamino, aminoalkyl, arylamino,            sulfonyl, alkylsulfonyl, arylsulfonyl, aminosulfonyl,            aminoalkyl, alkoxyalky, —COOH, C(O)OR⁵, —COR⁵, —SH, —SR⁶,            —OR⁶ and acyl;        -   L¹ and L² are the same or different and independently C₁-C₅            alkyl, which may be optionally substituted with one or more            substituents independently selected from the group            consisting of: halogen; ═O; ═S; —CN; —NO₂; —CF₃, —OCF₃,            alkyl, alkoxy, acylamino and alkylamino;        -   W is selected from the group consisting of a single bond,            —O—, —S—, —S(O)—, —S(O)₂—, —N(R⁹)—, —C(O)N(R⁹)—, —SO₂N(R⁹)—,            N(R⁹)C(O)—, N(R⁹)SO₂—, and —N(R⁹)—C(O)—N(R¹⁰)—;    -   c) L=Cy-(CH₂)m-W—    -   Wherein,        -   Cy is C₁-C₁₅ alkyl, aminoalkyl, heterocycloalkyl,            cycloalkyl, aryl, aryloxy or heteroaryl, any of which may be            optionally substituted one or more substituents            independently selected from the group consisting of:            halogen, ═O, ═S, —CN, —NO₂, —CF₃, —OCF₃, alkyl, alkenyl,            alkynyl, haloalkyl, haloalkenyl, haloalkynyl, heteroalkyl,            cycloalkyl, cycloalkenyl, heterocycloalkyl,            heterocycloalkenyl, aryl, heteroaryl, hydroxy, hydroxyalkyl,            alkoxy, alkoxyalkyl, alkoxyaryl, alkoxyheteroaryl,            alkenyloxy, alkynyloxy, cycloalkyloxy, cycloalkenyloxy,            heterocycloalkyloxy, heterocycloalkenyloxy, aryloxy,            heteroaryloxy, arylalkyl, heteroarylalkyl, arylalkyloxy,            amino, alkylamino, acylamino, aminoalkyl, arylamino,            sulfonyl, alkylsulfonyl, arylsulfonyl, aminosulfonyl,            aminoalkyl, alkoxyalky, —COOH, C(O)OR⁵, —COR⁵, —SH, —SR⁶,            —OR⁶ and acyl;        -   m is 0, 1, 2, 3, 4 or 5;        -   W is selected from the group consisting of a single bond,            —O—, —S—, —S(O)—, —S(O)₂—, —N(R⁹)—, —C(O)N(R⁹)—, —SO₂N(R⁹)—,            N(R⁹)C(O)—, N(R⁹)SO₂—, and —N(R⁹)—C(O)—N(R¹⁰)—;    -   d) L=L¹-W-L²        -   L¹ and L² are the same or different and independently            selected from C₁-C₅ alkyl, which may be optionally            substituted one or more substituents independently selected            from the group consisting of: halogen; ═O; ═S; —CN; —NO₂;            —CF₃, —OCF₃, alkyl, alkoxy, acylamino, alkylamino;        -   W is selected from the group consisting of a single bond,            —O—, —S—, —S(O)—, —S(O)₂—, —N(R⁹)—, —C(O)N(R⁹)—, —SO₂N(R⁹)—,            N(R⁹)C(O)—, N(R⁹)SO₂—, and —N(R⁹)—C(O)—N(R¹⁰)—;

R⁹ and R¹⁰ are the same or different and are independently selected fromH, C₁-C₆ alkyl, C₄-C₉ cycloalkyl, C₄-C₉ heterocycloalkyl, aryl,heteroaryl, arylalkyl and heteroarylalkyl; and acyl;

Z is a single bond or is selected from —CH₂—, —CH₂CH₂—, —CH═CH— andC₃-C₆ cycloalkyl, unsubstituted or substituted with one or moresubstituents independently selected from the group consisting of C₁-C₄alkyl; or a pharmaceutically acceptable salt thereof.

One suitable genus of hydroxamic compounds are those of formula Ia:

wherein

R¹ is selected from the group consisting of: H, alkyl, alkenyl, alkynyl,haloalkyl, haloalkenyl, heteroalkyl, cycloalkyl, cycloalkenyl,heterocycloalkyl, heterocycloalkenyl, aryl, heteroaryl, cycloalkylalkyl,heterocycloalkylalkyl, arylalkyl, heteroarylalkyl, arylalkenyl,cycloalkylheteroalkyl, arylheteroalkyl, heterocycloalkylheteroalkyl,heteroarylheteroalkyl, hydroxy, hydroxyalkyl, alkoxy, alkoxyalkyl,alkoxyaryl, alkenyloxy, alkynyloxy, cycloalkylkoxy, heterocycloalkyloxy,aryloxy, heteroaryloxy, arylalkyloxy, amino, alkylamino, aminoalkyl,acylamino, arylamino, phenoxy, benzyloxy, COOH, alkoxycarbonyl,alkylaminocarbonyl, sulfonyl, alkylsulfonyl, alkylsulfinyl,arylsulfonyl, arylsulfinyl, aminosulfonyl, SR⁶ and acyl, each of whichmay be unsubstituted or substituted with one or more substituentsindependently selected from the group consisting of: halogen, ═O, ═S,—CN, —NO₂, —CF₃, —OCF₃, alkyl, alkenyl, alkynyl, haloalkyl, haloalkenyl,haloalkynyl, heteroalkyl, cycloalkyl, cycloalkenyl, heterocycloalkyl,heterocycloalkenyl, aryl, heteroaryl, hydroxy, hydroxyalkyl, alkoxy,alkoxyalkyl, alkoxyaryl, alkoxyheteroaryl, alkenyloxy, alkynyloxy,cycloalkyloxy, cycloalkenyloxy, heterocycloalkyloxy,heterocycloalkenyloxy, aryloxy, heteroaryloxy, arylalkyl,heteroarylalkyl, arylalkyloxy, amino, alkylamino, acylamino, aminoalkyl,arylamino, sulfonyl, alkylsulfonyl, arylsulfonyl, arylsulfinyl,aminosulfonyl, aminoalkyl, alkoxyalky, —COOH, —C(O)OR⁵, —COR⁵, —SH,—SR⁶, —OR⁶ and acyl;

or R¹=L;

R² is selected from the group consisting of: H, halogen, alkyl, alkenyl,alkynyl, haloalkyl, haloalkenyl, heteroalkyl, cycloalkyl, cycloalkenyl,heterocycloalkyl, heterocycloalkenyl, aryl, heteroaryl, cycloalkylalkyl,heterocycloalkylalkyl, arylalkyl, heteroarylalkyl, arylalkenyl,cycloalkylheteroalkyl, heterocycloalkylheteroalkyl,heteroarylheteroalkyl, arylheteroalkyl hydroxy, hydroxyalkyl, alkoxy,alkoxyalkyl, alkoxyaryl, alkenyloxy, alkynyloxy, cycloalkylkoxy,heterocycloalkyloxy, aryloxy, heteroaryloxy, arylalkyloxy, amino,alkylamino, aminoalkyl, acylamino, arylamino, phenoxy, benzyloxy, COOH,alkoxycarbonyl, alkylaminocarbonyl, sulfonyl, alkylsulfonyl,alkylsulfinyl, arylsulfonyl, arylsulfinyl, aminosulfonyl, SR⁶ and acyl,each of which may be unsubstituted or substituted with one or moresubstituents independently selected from the group consisting of:halogen, ═O, ═S, —CN, —NO₂, —CF₃, —OCF₃, alkyl, alkenyl, alkynyl,haloalkyl, haloalkenyl, haloalkynyl, heteroalkyl, cycloalkyl,cycloalkenyl, heterocycloalkyl, heterocycloalkenyl, aryl, heteroaryl,hydroxy, hydroxyalkyl, alkoxy, alkoxyalkyl, alkoxyaryl,alkoxyheteroaryl, alkenyloxy, alkynyloxy, cycloalkyloxy,cycloalkenyloxy, heterocycloalkyloxy, heterocycloalkenyloxy, aryloxy,heteroaryloxy, arylalkyl, heteroarylalkyl, arylalkyloxy, amino,alkylamino, acylamino, aminoalkyl, arylamino, sulfonyl, alkylsulfonyl,arylsulfonyl, aminosulfonyl, aminoalkyl, alkoxyalky, —COOH, —COR⁵,—C(O)OR⁵, —SH, —SR⁶, —OR⁶ and acyl;

or R²=L;

R³ is selected from the group consisting of: H, C₁-C₆ alkyl, and acyl;or a metal ion selected from sodium, calcium, magnesium;

X and Y are the same or different and are independently selected fromthe group consisting of: H, halogen, —CN, —NO₂, —CF₃, —OCF₃, alkyl,alkenyl, alkynyl, haloalkyl, haloalkenyl, haloalkynyl, heteroalkyl,cycloalkyl, cycloalkenyl, heterocycloalkyl, heterocycloalkenyl, aryl,heteroaryl, hydroxy, hydroxyalkyl, alkoxy, alkoxyalkyl, alkoxyaryl,alkoxyheteroaryl, alkenyloxy, alkynyloxy, cycloalkyloxy,cycloalkenyloxy, heterocycloalkyloxy, heterocycloalkenyloxy, aryloxy,heteroaryloxy, arylalkyl, heteroarylalkyl, arylalkyloxy, amino,alkylamino, acylamino, aminoalkyl, arylamino, sulfonyl, alkylsulfonyl,arylsulfonyl, aminosulfonyl, aminoalkyl, alkoxyalky, —COOH, —C(O)OR⁵,—COR⁵, —SH, —SR⁶, —OR⁶, acyl and —NR⁷R⁸;

Each R⁵ is independently selected from the group consisting of: alkyl,alkenyl, alkynyl, haloalkyl, heteroalkyl, cycloalkyl, heterocycloalkyl,aryl, heteroaryl, cycloalkylalkyl, heterocycloalkylalkyl, arylalkyl,heteroarylalkyl and acyl;

Each R⁶ is independently selected from the group consisting of: alkyl,alkenyl, alkynyl, haloalkyl, heteroalkyl, cycloalkyl, heterocycloalkyl,aryl, heteroaryl, cycloalkylalkyl, heterocycloalkylalkyl, arylalkyl,heteroarylalkyl and acyl;

Each R⁷ and R⁸ are each independently selected from the group consistingof: alkyl, alkenyl, alkynyl, haloalkyl, heteroalkyl, cycloalkyl,heterocycloalkyl, aryl, heteroaryl, cycloalkylalkyl,heterocycloalkylalkyl, arylalkyl, heteroarylalkyl and acyl;

L is selected from the group consisting of:

-   -   a) L=Cy-L¹-W—    -   Wherein    -   Cy is C₁-C₁₅ alkyl, aminoalkyl, heterocycloalkyl, cycloalkyl,        aryl, aryloxy, or heteroaryl any of which may be optionally        substituted one or more substituents independently selected from        the group consisting of: halogen, ═O, ═S, —CN, —NO₂, —CF₃,        —OCF₃, alkyl, alkenyl, alkynyl, haloalkyl, haloalkenyl,        haloalkynyl, heteroalkyl, cycloalkyl, cycloalkenyl,        heterocycloalkyl, heterocycloalkenyl, aryl, heteroaryl, hydroxy,        hydroxyalkyl, alkoxy, alkoxyalkyl, alkoxyaryl, alkoxyheteroaryl,        alkenyloxy, alkynyloxy, cycloalkyloxy, cycloalkenyloxy,        heterocycloalkyloxy, heterocycloalkenyloxy, aryloxy,        heteroaryloxy, arylalkyl, heteroarylalkyl, arylalkyloxy, amino,        alkylamino, acylamino, aminoalkyl, arylamino, sulfonyl,        alkylsulfonyl, arylsulfonyl, aminosulfonyl, aminoalkyl,        alkoxyalky, —COOH, —C(O)OR⁵, —COR⁵, —SH, —SR⁶, —OR⁶, and acyl.        -   L¹ is selected from the group consisting of C₁-C₅ alkyl,            which may be optionally substituted with one or more            substituents independently selected from the group            consisting of: halogen; ═O; ═S; —CN; —NO₂; alkyl, alkoxy,            acylamino, and alkylamino;        -   W is selected from the group consisting of a single bond,            —O—, —S—, —S(O)—, —S(O)₂—, —N(R⁹)—, —C(O)N(R⁹)—, —SO₂N(R⁹)—,            N(R⁹)C(O)—, N(R⁹)SO₂—, and —N(R⁹)—C(O)—N(R¹⁰)—;    -   b) L=Cy-L¹-W-L²    -   Wherein,        -   Cy is C₁-C₁₅ alkyl, aminoalkyl, heterocycloalkyl,            cycloalkyl, aryl, aryloxy or heteroaryl, any of which may be            optionally substituted one or more substituents            independently selected from the group consisting of:            halogen, ═O, ═S, —CN, —NO₂, —CF₃, —OCF₃, alkyl, alkenyl,            alkynyl, haloalkyl, haloalkenyl, haloalkynyl, heteroalkyl,            cycloalkyl, cycloalkenyl, heterocycloalkyl,            heterocycloalkenyl, aryl, heteroaryl, hydroxy, hydroxyalkyl,            alkoxy, alkoxyalkyl, alkoxyaryl, alkoxyheteroaryl,            alkenyloxy, alkynyloxy, cycloalkyloxy, cycloalkenyloxy,            heterocycloalkyloxy, heterocycloalkenyloxy, aryloxy,            heteroaryloxy, arylalkyl, heteroarylalkyl, arylalkyloxy,            amino, alkylamino, acylamino, aminoalkyl, arylamino,            sulfonyl, alkylsulfonyl, arylsulfonyl, aminosulfonyl,            aminoalkyl, alkoxyalky, —COOH, C(O)OR⁵, —COR⁵, —SH, —SR⁶,            —OR⁶ and acyl;        -   L¹ and L² are the same or different and independently C₁-C₅            alkyl, which may be optionally substituted with one or more            substituents independently selected from the group            consisting of: halogen; ═O; ═S; —CN; —NO₂; —CF₃, —OCF₃,            alkyl, alkoxy, acylamino and alkylamino;        -   W is selected from the group consisting of a single bond,            —O—, —S—, —S(O)—, —S(O)₂—, —N(R⁹)—, —C(O)N(R⁹)—, —SO₂N(R⁹)—,            N(R⁹)C(O)—, N(R⁹)SO₂—, and —N(R⁹)—C(O)—N(R¹⁰)—;    -   c) L=Cy-(CH₂)m-W—    -   Wherein,        -   Cy is C₁-C₁₅ alkyl, aminoalkyl, heterocycloalkyl,            cycloalkyl, aryl, aryloxy or heteroaryl, any of which may be            optionally substituted one or more substituents            independently selected from the group consisting of halogen,            ═O, ═S, —CN, —NO₂, —CF₃, —OCF₃, alkyl, alkenyl, alkynyl,            haloalkyl, haloalkenyl, haloalkynyl, heteroalkyl,            cycloalkyl, cycloalkenyl, heterocycloalkyl,            heterocycloalkenyl, aryl, heteroaryl, hydroxy, hydroxyalkyl,            alkoxy, alkoxyalkyl, alkoxyaryl, alkoxyheteroaryl,            alkenyloxy, alkynyloxy, cycloalkyloxy, cycloalkenyloxy,            heterocycloalkyloxy, heterocycloalkenyloxy, aryloxy,            heteroaryloxy, arylalkyl, heteroarylalkyl, arylalkyloxy,            amino, alkylamino, acylamino, aminoalkyl, arylamino,            sulfonyl, alkylsulfonyl, arylsulfonyl, aminosulfonyl,            aminoalkyl, alkoxyalky, —COOH, C(O)OR⁵, —COR⁵, —SH, —SR⁶,            —OR⁶ and acyl;        -   m is 0, 1, 2, 3, 4 or 5;        -   W is selected from the group consisting of a single bond,            —O—, —S—, —S(O)—, —S(O)₂—, —N(R⁹)—, —C(O)N(R⁹)—, —SO₂N(R⁹)—,            N(R⁹)C(O)—, N(R⁹)SO₂—, and —N(R⁹)—C(O)—N(R¹⁰)—;    -   d) L=L¹-W-L²        -   L¹ and L² are the same or different and independently            selected from C₁-C₅ alkyl, which may be optionally            substituted one or more substituents independently selected            from the group consisting of: halogen; ═O; ═S; —CN; —NO₂;            —CF₃, —OCF₃, alkyl, alkoxy, acylamino, alkylamino;        -   W is selected from the group consisting of a single bond,            —O—, —S—, —S(O)—, —S(O)₂—, —N(R⁹)—, —C(O)N(R⁹)—, —SO₂N(R⁹)—,            N(R⁹)C(O)—, N(R⁹)SO₂—, and —N(R⁹)—C(O)—N(R¹⁰)—;

R⁹ and R¹⁰ are the same or different and are independently selected fromH, C₁-C₆ alkyl, C₄-C₉ cycloalkyl, C₄-C₉ heterocycloalkyl, aryl,heteroaryl, arylalkyl, heteroarylalkyl; and acyl;

Z is a single bond or is selected from —CH₂—, —CH₂CH₂—, —CH═CH—, C₃-C₆cycloalkyl, unsubstituted or substituted with one or more substituentsindependently selected from the group consisting of C₁-C₄ alkyl;

or a pharmaceutically acceptable salt thereof.

Another group of useful compounds are those of the formula Ib:

wherein

R¹ is selected from the group consisting of: H, alkyl, alkenyl, alkynyl,haloalkyl, haloalkenyl, heteroalkyl, cycloalkyl, cycloalkenyl,heterocycloalkyl, heterocycloalkenyl, aryl, heteroaryl, cycloalkylalkyl,heterocycloalkylalkyl, arylalkyl, heteroarylalkyl, arylalkenyl,cycloalkylheteroalkyl, arylheteroalkyl, heterocycloalkylheteroalkyl,heteroarylheteroalkyl, hydroxy, hydroxyalkyl, alkoxy, alkoxyalkyl,alkoxyaryl, alkenyloxy, alkynyloxy, cycloalkylkoxy, heterocycloalkyloxy,aryloxy, heteroaryloxy, arylalkyloxy, amino, alkylamino, aminoalkyl,acylamino, arylamino, phenoxy, benzyloxy, COOH, alkoxycarbonyl,alkylaminocarbonyl, sulfonyl, alkylsulfonyl, alkylsulfinyl,arylsulfonyl, arylsulfinyl, aminosulfonyl, SR⁶ and acyl, each of whichmay be unsubstituted or substituted with one or more substituentsindependently selected from the group consisting of: halogen, ═O, ═S,—CN, —NO₂, —CF₃, —OCF₃, alkyl, alkenyl, alkynyl, haloalkyl, haloalkenyl,haloalkynyl, heteroalkyl, cycloalkyl, cycloalkenyl, heterocycloalkyl,heterocycloalkenyl, aryl, heteroaryl, hydroxy, hydroxyalkyl, alkoxy,alkoxyalkyl, alkoxyaryl, alkoxyheteroaryl, alkenyloxy, alkynyloxy,cycloalkyloxy, cycloalkenyloxy, heterocycloalkyloxy,heterocycloalkenyloxy, aryloxy, heteroaryloxy, arylalkyl,heteroarylalkyl, arylalkyloxy, amino, alkylamino, acylamino, aminoalkyl,arylamino, sulfonyl, alkylsulfonyl, arylsulfonyl, arylsulfinyl,aminosulfonyl, aminoalkyl, alkoxyalky, —COOH, —C(O)OR⁵, —COR⁵, —SH,—SR⁶, —OR⁶ and acyl;

or R¹=L;

R² is selected from the group consisting of: H, halogen, alkyl, alkenyl,alkynyl, haloalkyl, haloalkenyl, heteroalkyl, cycloalkyl, cycloalkenyl,heterocycloalkyl, heterocycloalkenyl, aryl, heteroaryl, cycloalkylalkyl,heterocycloalkylalkyl, arylalkyl, heteroarylalkyl, arylalkenyl,cycloalkylheteroalkyl, heterocycloalkylheteroalkyl,heteroarylheteroalkyl, arylheteroalkyl hydroxy, hydroxyalkyl, alkoxy,alkoxyalkyl, alkoxyaryl, alkenyloxy, alkynyloxy, cycloalkylkoxy,heterocycloalkyloxy, aryloxy, heteroaryloxy, arylalkyloxy, amino,alkylamino, aminoalkyl, acylamino, arylamino, phenoxy, benzyloxy, COOH,alkoxycarbonyl, alkylaminocarbonyl, sulfonyl, alkylsulfonyl,alkylsulfinyl, arylsulfonyl, arylsulfinyl, aminosulfonyl, SR⁶ and acyl,each of which may be unsubstituted or substituted with one or moresubstituents independently selected from the group consisting of:halogen, ═O, ═S, —CN. —NO₂, —CF₃, —OCF₃, alkyl, alkenyl, alkynyl,haloalkyl, haloalkenyl, haloalkynyl, heteroalkyl, cycloalkyl,cycloalkenyl, heterocycloalkyl, heterocycloalkenyl, aryl, heteroaryl,hydroxy, hydroxyalkyl, alkoxy, alkoxyalkyl, alkoxyaryl,alkoxyheteroaryl, alkenyloxy, alkynyloxy, cycloalkyloxy,cycloalkenyloxy, heterocycloalkyloxy, heterocycloalkenyloxy, aryloxy,heteroaryloxy, arylalkyl, heteroarylalkyl, arylalkyloxy, amino,alkylamino, acylamino, aminoalkyl, arylamino, sulfonyl, alkylsulfonyl,arylsulfonyl, aminosulfonyl, aminoalkyl, alkoxyalky, —COOH, —COR⁵,—C(O)OR⁵, —SH, —SR⁶, —OR⁶ and acyl;

or R²=L;

X and Y are the same or different and are independently selected fromthe group consisting of: H, halogen, —CN, —NO₂, —CF₃, —OCF₃, alkyl,alkenyl, alkynyl, haloalkyl, haloalkenyl, haloalkynyl, heteroalkyl,cycloalkyl, cycloalkenyl, heterocycloalkyl, heterocycloalkenyl, aryl,heteroaryl, hydroxy, hydroxyalkyl, alkoxy, alkoxyalkyl, alkoxyaryl,alkoxyheteroaryl, alkenyloxy, alkynyloxy, cycloalkyloxy,cycloalkenyloxy, heterocycloalkyloxy, heterocycloalkenyloxy, aryloxy,heteroaryloxy, arylalkyl, heteroarylalkyl, arylalkyloxy, amino,alkylamino, acylamino, aminoalkyl, arylamino, sulfonyl, alkylsulfonyl,arylsulfonyl, aminosulfonyl, aminoalkyl, alkoxyalky, —COOH, —C(O)OR⁵,—COR⁵, —SH, —SR⁶, acyl and —NR⁷R⁸;

Each R⁵ is independently selected from the group consisting of: alkyl,alkenyl, alkynyl, haloalkyl, heteroalkyl, cycloalkyl, heterocycloalkyl,aryl, heteroaryl, cycloalkylalkyl, heterocycloalkylalkyl, arylalkyl,heteroarylalkyl and acyl;

Each R⁶ is independently selected from the group consisting of: alkyl,alkenyl, alkynyl, haloalkyl, heteroalkyl, cycloalkyl, heterocycloalkyl,aryl, heteroaryl, cycloalkylalkyl, heterocycloalkylalkyl, arylalkyl,heteroarylalkyl and acyl;

Each R⁷ and R⁸ are each independently selected from the group consistingof: H, alkyl, alkenyl, alkynyl, haloalkyl, heteroalkyl, cycloalkyl,heterocycloalkyl, aryl, heteroaryl, cycloalkylalkyl,heterocycloalkylalkyl, arylalkyl, heteroarylalkyl and acyl;

L is selected from the group consisting of:

-   -   a) L=Cy-L¹-W—    -   Wherein        -   Cy is C₁-C₁₅ alkyl, aminoalkyl, heterocycloalkyl,            cycloalkyl, aryl, aryloxy or heteroaryl, any of which may be            optionally substituted one or more substituents            independently selected from the group consisting of:            halogen, ═O, ═S, —CN, —NO₂, —CF₃, —OCF₃, alkyl, alkenyl,            alkynyl, haloalkyl, haloalkenyl, haloalkynyl, heteroalkyl,            cycloalkyl, cycloalkenyl, heterocycloalkyl,            heterocycloalkenyl, aryl, heteroaryl, hydroxy, hydroxyalkyl,            alkoxy, alkoxyalkyl, alkoxyaryl, alkoxyheteroaryl,            alkenyloxy, alkynyloxy, cycloalkyloxy, cycloalkenyloxy,            heterocycloalkyloxy, heterocycloalkenyloxy, aryloxy,            heteroaryloxy, arylalkyl, heteroarylalkyl, arylalkyloxy,            amino, alkylamino, acylamino, aminoalkyl, arylamino,            sulfonyl, alkylsulfony), arylsulfonyl, aminosulfonyl,            aminoalkyl, alkoxyalky, —COOH, —C(O)OR⁵, —COR⁵, —SH, —SR⁶,            —OR⁶ and acyl.        -   L² is selected from the group consisting of C₁-C₅ alkyl,            which may be optionally substituted with one or more            substituents independently selected from the group            consisting of: halogen; ═O; ═S; —CN; —NO₂; alkyl, alkoxy,            acylamino, and alkylamino;        -   W is selected from the group consisting of a single bond,            —O—, —S—, —S(O)—, —S(O)₂—, —N(R⁹)—, —C(O)N(R⁹)—, —SO₂N(R⁹)—,            N(R⁹)C(O)—, N(R⁹)SO₂—, and —N(R⁹)—C(O)—N(R¹⁰)—;    -   b) L=Cy-L¹-W-L²    -   Wherein,        -   Cy is C₁-C₁₅ alkyl, aminoalkyl, heterocycloalkyl,            cycloalkyl, aryl, aryloxy or heteroaryl, any of which may be            optionally substituted one or more substituents            independently selected from the group consisting of:            halogen, ═O, ═S, —CN, —NO₂, —CF₃, —OCF₃, alkyl, alkenyl,            alkynyl, haloalkyl, haloalkenyl, haloalkynyl, heteroalkyl,            cycloalkyl, cycloalkenyl, heterocycloalkyl,            heterocycloalkenyl, aryl, heteroaryl, hydroxy, hydroxyalkyl,            alkoxy, alkoxyalkyl, alkoxyaryl, alkoxyheteroaryl,            alkenyloxy, alkynyloxy, cycloalkyloxy, cycloalkenyloxy,            heterocycloalkyloxy, heterocycloalkenyloxy, aryloxy,            heteroaryloxy, arylalkyl, heteroarylalkyl, arylalkyloxy,            amino, alkylamino, acylamino, aminoalkyl, arylamino,            sulfonyl, alkylsulfonyl, arylsulfonyl, aminosulfonyl,            aminoalkyl, alkoxyalky, —COOH, C(O)OR⁵, —COR⁵, —SH, —SR⁶,            —OR⁶ and acyl;        -   L¹ and L² are the same or different and independently C₁-C₅            alkyl, which may be optionally substituted with one or more            substituents independently selected from the group            consisting of: halogen; ═O; ═S; —CN; —NO₂; —CF₃, —OCF₃,            alkyl, alkoxy, acylamino and alkylamino;        -   W is selected from the group consisting of a single bond,            —O—, —S—, —S(O)—, —S(O)₂—, —N(R′)—, —C(O)N(R⁹)—, —SO₂N(R⁹)—,            N(R⁹)C(O)—, N(R⁹)SO₂—, and —N(R⁹)—C(O)—N(R¹⁰)—;    -   c) L=Cy-(CH₂)m-W—    -   Wherein,        -   Cy is C₁-C₁₅ alkyl, aminoalkyl, heterocycloalkyl,            cycloalkyl, aryl, aryloxy or heteroaryl, any of which may be            optionally substituted one or more substituents            independently selected from the group consisting of:            halogen, ═O, ═S, —CN, —NO₂, —CF₃, —OCF₃, alkyl, alkenyl,            alkynyl, haloalkyl, haloalkenyl, haloalkynyl, heteroalkyl,            cycloalkyl, cycloalkenyl, heterocycloalkyl,            heterocycloalkenyl, aryl, heteroaryl, hydroxy, hydroxyalkyl,            alkoxy, alkoxyalkyl, alkoxyaryl, alkoxyheteroaryl,            alkenyloxy, alkynyloxy, cycloalkyloxy, cycloalkenyloxy,            heterocycloalkyloxy, heterocycloalkenyloxy, aryloxy,            heteroaryloxy, arylalkyl, heteroarylalkyl, arylalkyloxy,            amino, alkylamino, acylamino, aminoalkyl, arylamino,            sulfonyl, alkylsulfonyl, arylsulfonyl, aminosulfonyl,            aminoalkyl, alkoxyalky, —COOH, C(O)OR⁵, —COR⁵, —SH, —SR⁶,            —OR⁶ and acyl        -   m is 0, 1, 2, 3, 4 or 5;        -   W is selected from the group consisting of a single bond,            —O—, —S—, —S(O)—, —S(O)₂—, —N(R⁹)—, —C(O)N(R⁹)—, —SO₂N(R⁹)—,            N(R⁹)C(O)—, N(R⁹)SO₂—, and —N(R⁹)—C(O)—N(R¹⁰)—;    -   d) L=L-W-L²        -   L¹ and L² are the same or different and independently            selected from C₁-C₅ alkyl, which may be optionally            substituted one or more substituents independently selected            from the group consisting of: halogen; ═O; ═S; —CN; —NO₂;            —CF₃, —OCF₃, alkyl, alkoxy, acylamino, alkylamino;        -   W is selected from the group consisting of a single bond,            —O—, —S—, —S(O)—, —S(O)₂—, —N(R⁹)—, —C(O)N(R⁹)—, —SO₂N(R⁹)—,            N(R⁹)C(O)—, N(R⁹)SO₂—, and —N(R⁹)—C(O)—N(R¹⁰)—;

R⁹ and R¹⁰ are the same or different and are independently selected fromH, C₁-C₆ alkyl, C₄-C₉ cycloalkyl, C₄-C₉ heterocycloalkyl, aryl,heteroaryl, arylalkyl, heteroarylalkyl; and acyl;

Z is a single bond or is selected from —CH₂—, —CH₂CH₂—, —CH═CH—, C₃-C₆cycloalkyl, unsubstituted or substituted with one or more substituentsindependently selected from the group consisting of C₁-C₄ alkyl;

or a pharmaceutically acceptable salt thereof.

As with any group of structurally related compounds which possess aparticular utility, certain groups are preferred for the compounds ofthe Formula (I), (Ia) and (Ib) in their end use application.

In certain preferred embodiments R¹ is selected from the groupconsisting of C₁-C₁₀ alkyl, alkenyl, heteroalkyl, haloalkyl, alkynyl,aryl, cycloalkyl, heterocycloalkyl, heteroaryl, C₄-C₉heterocycloalkylalkyl, cycloalkylalkyl, arylalkyl, and heteroarylalkyleach of which may be substituted as previously stated.

In another embodiment it is preferred that R¹ is selected from the groupconsisting of H, hydroxyalkyl, alkyl, arylalkyl, heteroarylalkyl,alkoxyalkyl, aminoalkyl, and heterocycloalkyl each of which may besubstituted as previously stated.

In another embodiment it is preferred that R¹ is selected from the groupconsisting of H, hydroxyalkyl, alkyl, alkoxyalkyl, and aminoalkyl eachof which may be substituted as previously stated.

In another embodiment it is preferred that if R¹ is alkyl or heteroalkylthen it is not substituted by a cycloalkyl, aryl, heteroaryl, orheterocycloalkyl.

Particularly preferred values of R¹ are:

H; methyl; (pyridin-2-yl)methyl; (pyridin-3-yl)methyl; ethyl;2-hydroxy-ethyl; 2-(pyridin-2-yl)ethyl; 2-(pyridin-3-yl)ethyl;2-phenyl-ethyl; 2-carboxy-ethyl; 2-(morpholin-4-yl)-ethyl;2-(piperidin-1-yl)-ethyl; 2-(pyrollidin-1-yl)-ethyl;2-diethylamino-ethyl; propyl; 2,3-di-hydroxy-propyl; 3-hydroxy-propyl;3-methoxy-propyl; 3-isopropoxy-propyl; 2,2-dimethyl-propyl;3-dimethylamino-propyl; 3-dimethylamino-2,2-dimethyl-propyl;3-(2-oxo-pyrollidin-1-yl)-propyl; 3-(morpholin-4-yl)-propyl;3-(imadazol-1-yl)-propyl; 3-(4-methyl-piperidin-1-yl)-propyl;3-(pyrollidin-1-yl)-propyl; 4-dimethylamino-butyl; 5-hydroxy-pentyl;allyl; benzyl; and 3,4,5-trimethoxybenzyl.

In certain preferred embodiments R² is selected from the groupconsisting of H, Halogen, C₁-C₁₀ alkyl, alkenyl, heteroalkyl, haloalkyl,alkynyl, aryl, cycloalkyl, heterocycloalkyl, heteroaryl, C₄-C₉heterocycloalkylalkyl, cycloalkylalkyl, arylalkyl, and heteroarylalkyleach of which may be substituted as previously stated.

In another embodiment it is preferred that R² is selected from the groupconsisting of H, alkyl, arylalkyl, aryl, heteroaryl, heteroalkyl,cycloalkyl, and L, each of which may be substituted as previouslystated.

In another embodiment it is preferred that R² is selected from the groupconsisting of H, hydroxyalkyl, alkyl, alkoxyalkyl, and aminoalkyl eachof which may be substituted as previously stated.

In another embodiment it is preferred that if R² is alkyl or heteroalkylthen it is not substituted by a cycloalkyl, aryl, heteroaryl, orheterocycloalkyl.

Particularly preferred values of R² are: H; methyl; benzylamino-methyl;dibenzylamino-methyl; [2-(4-fluoro-phenyl)-acetylamino]-methyl;[2-(4-methoxy-phenyl)-acetylamino]-methyl; 4-methoxy-benzylamino-methyl;benzyloxy-methyl; phenylacetylamino-methyl; 1-amino-2-phenyl-ethyl;2-benzylamino-ethyl; 2-(3-methoxy-phenyl)-ethyl; 2-(pyridin-3-yl)ethyl;2-(2-phenoxyacetylamino)-ethyl; 2-benzenesulphonylamino-ethyl;2-phenyl-ethyl; isopropyl; 2-phenyl-propyl; 3-phenyl-propyl;3-phenoxy-propyl; 3-(1H-indol-3-yl)-propyl; 4-methoxy-phenyl;4-fluoro-phenyl; 4-benzyloxy-3-methoxy-phenyl; isobutyl; cyclohexyl;octyl; benzyl; pyridin-2-yl; pyridin-4-yl; thiophen-3-yl;benzylsulfanyl, and 2-phenylmethansulfanyl.

If R¹ or R² are substituted particularly preferred substituents areselected from the group consisting of: halogen, ═O, ═S, —CN, —NO₂,alkyl, alkenyl, heteroalkyl, haloalkyl, alkynyl, aryl, cycloalkyl,heterocycloalkyl, heteroaryl, hydroxy, hydroxyalkyl, alkoxy, alkylamino,aminoalkyl, acylamino, phenoxy, alkoxyalkyl, benzyloxy, alkylsulfonyl,arylsulfonyl, aminosulfonyl, —C(O)OR⁵, COOH, SH, and acyl.

X and Y may be the same or different and are preferably H, halogen,C₁-C₄ alkyl, —CF₃, —NO₂, —C(O)R⁵, —OR⁶, —SR⁶, —CN and NR⁷R⁸.

X is most preferably H;Y is most preferably H;

X and Y are most preferably at the 4 and 7 positions of the aromaticring.

R₃ is preferably H, C₁-C₆ alkyl, or acyl, more preferably H or C₁-C₄alkyl, most preferably H;

R₄ is preferably H or C₁-C₄ alkyl, most preferably H;R₅ is preferably C₁-C₄ alkyl, heteroalkyl, or acyl, most preferablymethyl;R₆ is preferably C₁-C₄ alkyl, heteroalkyl or acyl, most preferably C₁-C₄alkyl;R₇ and Re are preferably selected from the group consisting of H, C₁-C₆alkyl, C₄-C₉cycloalkyl, C₄-C₉heterocycloalkyl, aryl, heteroaryl,arylalkyl, and heteroarylalkyl

The Z moiety is preferably a group of formula —CH═CH—. The moiety ispreferably in the “E” configuration and is preferably at the 5 or 6position, most preferably the 5 position.

In addition to compounds of Formula I, the embodiments disclosed arealso directed to pharmaceutically acceptable salts, pharmaceuticallyacceptable prodrugs, and pharmaceutically active metabolites of suchcompounds, and pharmaceutically acceptable salts of such metabolites.Such compounds, salts, prodrugs and metabolites are at timescollectively referred to herein as “HDAC inhibiting agents” or “HDACinhibitors”.

The invention also relates to pharmaceutical compositions including acompound of the invention with a pharmaceutically acceptable carrier,diluent or excipient.

In yet a further aspect the present invention provides a method oftreatment of a disorder caused by, associated with or accompanied bydisruptions of cell proliferation and/or angiogenesis includingadministration of a therapeutically effective amount of a compound offormula (I).

The method preferably includes administration of a compound of formula(Ia), more preferably a compound of formula (Ib) as described herein.

The disorder is preferably selected from the group consisting of but notlimited to cancer, inflammatory diseases/immune system disorders,angiofibroma, cardiovascular diseases (e.g. restenosis,arteriosclerosis), fibrotic diseases (e.g. liver fibrosis), diabetes,autoimmune diseases, chronic and acute neurodegenerative disease likedisruptions of nerval tissue, Huntington's disease and infectiousdiseases like fungal, bacterial and viral infections. In anotherembodiment the disorder is a proliferative disorder. The proliferativedisorder is preferably cancer.

The invention also provides agents for the treatment of a disordercaused by, associated with or accompanied by disruptions of cellproliferation and/or angiogenesis including a compound of formula (I) asdisclosed herein. The agent is preferably an anti-cancer agent.

The agent preferably contains a compound of formula (Ia), morepreferably a compound of formula (Ib).

The invention also relates to the use of compounds of formula (I) in thepreparation of a medicament for the treatment of a disorder caused by,associated with or accompanied by disruptions of cell proliferationand/or angiogenesis. The disorder is preferably a proliferativedisorder, most preferably a cancer.

The compounds of the present invention surprisingly show low toxicity,together with a potent anti-proliferative activity.

In yet a further embodiment the invention provides a method of treatmentof a disorder, disease or condition that can be treated by theinhibition of histone deacetylase including administration of atherapeutically effective amount of a compound of formula (1).

The method preferably includes administration of a compound of formula(Ia), more preferably a compound of formula (Ib) as described herein.

The disorder is preferably selected the group consisting of but notlimited to Anti-proliferative disorders (e.g. cancers);Neurodegenerative diseases including Huntington's Disease, Polyglutaminedisease, Parkinson's Disease, Alzheimer's Disease, Seizures,Striatonigral degeneration, Progressive supranuclear palsy, Torsiondystonia, Spasmodic torticollis and dyskinesis, Familial tremor, Gillesde la Tourette syndrome, Diffuse Lewy body disease, Progressivesupranuclear palsy, Pick's disease, Intracerebral haemorrhage Primarylateral sclerosis, Spinal muscular atrophy, Amyotrophic lateralsclerosis, Hypertrophic interstitial polyneuropathy, Retinitispigmentosa, Hereditary optic atrophy, Hereditary spastic paraplegia,Progressive ataxia and Shy-Drager syndrome; Metabolic diseases includingType 2 diabetes; Degenerative Diseases of the Eye including Glaucoma,Age-related macular degeneration, Rubeotic glaucoma; Inflammatorydiseases and/or Immune system disorders including Rheumatoid Arthritis(RA), Osteoarthritis, Juvenile chronic arthritis, Graft versus Hostdisease, Psoriasis, Asthma, Spondyloarthropathy, psoriasis, Crohn'sDisease, inflammatory bowel disease, Colitis Ulcerosa, Alcoholichepatitis, Diabetes, Sjoegrens's syndrome, Multiple Sclerosis,Ankylosing spondylitis, Membranous glomerulopathy, Discogenic pain,Systemic Lupus Erythematosus; Disease involving angiogenesis includingcancer, psoriasis, rheumatoid arthritis; Psychological disordersincluding bipolar disease, schizophrenia, depression and dementia;Cardiovascular Diseases including Heart failure, restenosis andarteriosclerosis; Fibrotic diseases including liver fibrosis, cysticfibrosis and angiofibroma; infectious diseases including Fungalinfections, such as Candida Albicans, Bacterial infections, Viralinfections, such as Herpes Simplex, Protozoal infections, such asMalaria, Leishmania infection, Trypanosoma brucei infection,Toxoplasmosis and coccidiosis and Haematopoietic disorders includingthalassemia, anemia and sickle cell anemia.

The invention also provides agents for the treatment of a disorder,disease or condition that can be treated by the inhibition of histonedeacetylase including a compound of formula (I) as disclosed herein. Theagent is preferably an anti-cancer agent.

The invention also relates to the use of compounds of formula (I) in thepreparation of a medicament for the treatment of a disorder, disease orcondition that can be treated by the inhibition of histone deacetylase.

The invention also provides a method for inhibiting cell proliferationincluding administration of an effective amount of a compound accordingto formula (I).

In yet an even further aspect the invention provides a method oftreatment of a neurodegenerative disorder in a patient includingadministration of a therapeutically effective amount of a compound offormula (I). The method preferably includes administration of a compoundof formula (Ia), more preferably a compound of formula (Ib) as describedherein. The neurodegenerative disorder is preferably Huntington'sDisease.

The invention also provides agents for the treatment ofneurodegenerative disorder including a compound of formula (I) asdisclosed herein. The agent is preferably anti-Huntington's diseaseagent.

The invention also relates to the use of compounds of formula (I) in thepreparation of a medicament for the treatment of a neurodegenerativedisorder. The neurodegenerative disorder is preferably Huntington'sDisease.

In yet an even further aspect the invention provides a method oftreatment of an inflammatory disease and/or immune system disorder in apatient including administration of a therapeutically effective amountof a compound of formula (1). The method preferably includesadministration of a compound of formula (Ia), more preferably a compoundof formula (Ib) as described herein. In one embodiment the inflammatorydisease and/or immune system disorder is rheumatoid arthritis. Inanother embodiment the inflammatory disease and/or immune systemdisorder is Systemic Lupus Erythematosus.

The invention also provides agents for the treatment of inflammatorydisease and/or immune system disorder including a compound of formula(I) as disclosed herein.

The invention also relates to the use of compounds of formula (I) in thepreparation of a medicament for the treatment of inflammatory diseaseand/or immune system disorder. In one embodiment the inflammatorydisease and/or immune system disorder is rheumatoid arthritis. Inanother embodiment the inflammatory disease and/or immune systemdisorder is Systemic Lupus Erythematosus.

To monitor the efficacy of such compounds the invention describes amethod suitable to detect and quantify levels of acetylated histone insamples from human or animal species such as tumor tissue, brain, andblood. The method is based on an enzyme-linked immunosorbant assay(ELISA) and may be used for the quantification of acetylated histones incellular extracts or samples from human or animal species such as tumortissue, brain, and blood. Preferable over conventional systems, theELISA allows high throughput, quantitative determinations of theconcentration of acetylated histones as measurement for the efficacy ofthe drug treatment or the potency of the drug in a respective biologicaltest system. For a general review of conventional ELISA techniques,please refer to Crowther J R (1995) ELISA theory and practice in Methodin molecular biology vol. 42, Humana.

In yet an even further aspect the invention provides a method formeasuring an acetylated histone concentration in a biological sampleusing an enzyme-linked immunosorbant assay, the enzyme-linkedimmunosorbant assay including a combination of a primary captureantibody, or a portion thereof, and secondary detection antibody, or aportion thereof.

The primary capture antibody is preferably selected from the groupconsisting of: an anti-H3 monoclonal antibody, an anti-acetylated H3polyclonal antibody, a goat anti-H3 polyclonal antibody, a goatanti-acetylated H3 polyclonal antibody and a combination thereof. Thesecondary detection antibody is preferably selected from the groupconsisting of: an anti-H3 monoclonal antibody, an anti-acetylated H3polyclonal antibody, a goat anti-H3 polyclonal antibody, a goatanti-acetylated H3 polyclonal antibody and a combination thereof.

In a particularly preferred embodiment the primary capture antibody is amouse anti-H3 monoclonal antibody and the secondary detection antibodyis a rat anti-acetylated H3 polyclonal antibody.

The invention also provides a method for identifying the pharmacologicaleffect of a histone deacetylase inhibitor in a cell, the methodincluding the steps of:

-   -   a) providing a cell that has been treated with a histone        deacetylase inhibitor;    -   b) measuring the acetylated histone concentration in the cell by        a method according to any one of claims 32 to 35; and    -   c) comparing the acetylated histone concentration with the        acetylated histone concentration of a control sample.

In a preferred embodiment the control sample is derived from a cell thathas not been treated with a histone deacetylase inhibitor. In anotherpreferred embodiment the cell is a tumour cell.

The histone deacetylase inhibitor preferably includes a compound offormula (1).

The invention also provides a method for identifying the pharmacologicaleffect of a histone deacetylase inhibitor in a subject, the methodincluding the steps of:

-   -   a) obtaining a biological sample from a subject that has been        treated with a histone deacetylase inhibitor;    -   b) measuring the acetylated histone concentration in the        biological sample by a method according to the invention as        described above; and    -   c) comparing the acetylated histone concentration with the        acetylated histone concentration of a control sample.

The control sample is preferably a biological sample derived from asubject that has not been treated with a histone deacetylase inhibitor.

In the methods of the invention the biological sample is preferablyselected from the group consisting of tissue, blood, serum, plasma,urine, saliva and a combination thereof.

DETAILED DESCRIPTION OF THE EMBODIMENTS

There are disclosed hydroxamate compounds, for example benzimidazolescontaining hydroxamic acid in one of the substituents, that may beinhibitors of deacetylases, including but not limited to inhibitors ofhistone deacetylases. The hydroxamate compounds may be suitable forprevention or treatment of a disorder caused by, associated with oraccompanied by disruptions of cell proliferation and/or angiogenesiswhen used either alone or together with a pharmaceutically acceptablecarrier, diluent or excipient. An example of such a disorder is cancer.

As used herein the term ‘cancer’ is a general term intended to encompassthe vast number of conditions that are characterised by uncontrolledabnormal growth of cells.

It is anticipated that the compounds of the invention will be useful intreating various cancers including but not limited to bone cancersincluding Ewing's sarcoma, osteosarcoma, chondrosarcoma and the like,brain and CNS tumours including acoustic neuroma, neuroblastomas, gliomaand other brain tumours, spinal cord tumours, breast cancers, colorectalcancers, advanced colorectal adenocarcinomas, endocrine cancersincluding adenocortical carcinoma, pancreatic cancer, pituitary cancer,thyroid cancer, parathyroid cancer, thymus cancer, multiple endocrineneoplasma, gastrointestinal cancers including stomach cancer, esophagealcancer, small intestine cancer, Liver cancer, extra hepatic bile ductcancer, gastrointestinal carcinoid tumour, gall bladder cancer,genitourinary cancers including testicular cancer, penile cancer,prostrate cancer, gynaecological cancers including cervical cancer,ovarian cancer, vaginal cancer, uterus/endometrium cancer, vulva cancer,gestational trophoblastic cancer, fallopian tube cancer, uterinesarcoma, head and neck cancers including oral cavity cancer, lip cancer,salivary gland cancer, larynx cancer, hypopharynx cancer, orthopharynxcancer, nasal cancer, paranasal cancer, nasopharynx cancer, leukemiasincluding childhood leukemia, acute lymphocytic leukemia, acute myeloidleukemia, chronic lymphocytic leukemia, chronic myeloid leukemia, hairycell leukemia, acute promyelocytic leukemia, plasma cell leukemia,myelomas, haematological disorders including myelodysplastic syndromes,myeloproliferative disorders, aplastic anemia, Fanconi anemia,Waldenstroms Macroglobulinemia, lung cancers including small cell lungcancer, non-small cell lung cancer, lymphomas including Hodgkin'sdisease, non-Hodgkin's lymphoma, cutaneous T-cell lymphoma, peripheralT-cell lymphoma, AIDS related Lymphoma, eye cancers includingretinoblastoma, intraocular melanoma, skin cancers including melanoma,non-melanoma skin cancer, merkel cell cancer, soft tissue sarcomas suchas childhood soft tissue sarcoma, adult soft tissue sarcoma, Kaposi'ssarcoma, urinary system cancers including kidney cancer, Wilms tumour,bladder cancer, urethral cancer, and transitional cell cancer.

Preferred cancers that may be treated by the compounds of the presentinvention are breast cancer, lung cancer, ovarian cancer, prostatecancer, head and neck cancer, renal, gastic and brain cancer.

Preferred cancers that may be treated by compounds of the presentinventions are cutaneous T-cell lymphoma (CTCL) and peripheral T-celllymphoma.

Preferred cancers that may be treated by compounds of the presentinvention include solid tumors and hematologic malignancies.

The compounds may also be used in the treatment of a disorder involving,relating to or, associated with dysregulation of histone deacetylase(HDAC).

There are a number of disorders that have been implicated by or known tobe mediated at least in part by HDAC activity, where HDAC activity isknown to play a role in triggering disease onset, or whose symptoms areknown or have been shown to be alleviated by HDAC inhibitors. Disordersof this type that would be expected to be amenable to treatment with thecompounds of the invention include the following but not limited to:

Anti-proliferative disorders (e.g. cancers); Neurodegenerative diseasesincluding Huntington's Disease, Polyglutamine disease, Parkinson'sDisease, Alzheimer's Disease, Seizures, Striatonigral degeneration,Progressive supranuclear palsy, Torsion dystonia, Spasmodic torticollisand dyskinesis, Familial tremor, Gilles de la Tourette syndrome, DiffuseLewy body disease, Progressive supranuclear palsy, Pick's disease,intracerebreal haemorrphage, Primary lateral sclerosis, Spinal muscularatrophy, Amyotrophic lateral sclerosis, Hypertrophic interstitialpolyneuropathy, Retinitis pigmentosa, Hereditary optic atrophy,Hereditary spastic paraplegia, Progressive ataxia and Shy-Dragersyndrome; Metabolic diseases including Type 2 diabetes; DegenerativeDiseases of the Eye including Glaucoma, Age-related maculardegeneration, Rubeotic glaucoma; Inflammatory diseases and/or Immunesystem disorders including Rheumatoid Arthritis (RA), Osteoarthritis,Juvenile chronic arthritis, Graft versus Host disease, Psoriasis,Asthma, Spondyloarthropathy, psoriasis, Crohn's Disease, inflammatorybowel disease Colitis Ulcerosa, Alcoholic hepatitis, Diabetes,Sjoegrens's syndrome, Multiple Sclerosis, Ankylosing spondylitis,Membranous glomerulopathy, Discogenic pain, Systemic LupusErythematosus; Disease involving angiogenesis including cancer,psoriasis, rheumatoid arthritis; Psychological disorders includingbipolar disease, schizophrenia, mainia, depression and dementia;Cardiovascular Diseases including heart failure, restenosis andarteriosclerosis; Fibrotic diseases including liver fibrosis, cysticfibrosis and angiofibroma; Infectious diseases including Fungalinfections, such as Candida Albicans, Bacterial infections, Viralinfections, such as Herpes Simplex, Protozoal infections, such asMalaria, Leishmania infection, Trypanosoma brucei infection,Toxoplasmosis and coccidiosis and Haematopoietic disorders includingthaiassemia, anemia and sickle cell anemia.

The hydroxamate compounds of the present invention have the followingstructure (I):

wherein

R¹ is selected from the group consisting of: H, alkyl, alkenyl, alkynyl,haloalkyl, haloalkenyl, heteroalkyl, cycloalkyl, cycloalkenyl,heterocycloalkyl, heterocycloalkenyl, aryl, heteroaryl, cycloalkylalkyl,heterocycloalkylalkyl, arylalkyl, heteroarylalkyl, arylalkenyl,cycloalkylheteroalkyl, arylheteroalkyl, heterocycloalkylheteroalkyl,heteroarylheteroalkyl, hydroxy, hydroxyalkyl, alkoxy, alkoxyalkyl,alkoxyaryl, alkenyloxy, alkynyloxy, cycloalkylkoxy, heterocycloalkyloxy,aryloxy, heteroaryloxy, arylalkyloxy, amino, alkylamino, aminoalkyl,acylamino, arylamino, phenoxy, benzyloxy, COOH, alkoxycarbonyl,alkylaminocarbonyl, sulfonyl, alkylsulfonyl, alkylsulfinyl,arylsulfonyl, arylsulfinyl, aminosulfonyl, SR⁶ and acyl, each of whichmay be unsubstituted or substituted with one or more substituentsindependently selected from the group consisting of: halogen, ═O, ═S,—CN, —NO₂, —CF₃, —OCF₃, alkyl, alkenyl, alkynyl, haloalkyl, haloalkenyl,haloalkynyl, heteroalkyl, cycloalkyl, cycloalkenyl, heterocycloalkyl,heterocycloalkenyl, aryl, heteroaryl, hydroxy, hydroxyalkyl, alkoxy,alkoxyalkyl, alkoxyaryl, alkoxyheteroaryl, alkenyloxy, alkynyloxy,cycloalkyloxy, cycloalkenyloxy, heterocycloalkyloxy,heterocycloalkenyloxy, aryloxy, heteroaryloxy, arylalkyl,heteroarylalkyl, arylalkyloxy, amino, alkylamino, acylamino, aminoalkyl,arylamino, sulfonyl, alkylsulfonyl, arylsulfonyl, arylsulfinyl,aminosulfonyl, aminoalkyl, alkoxyalky, —COOH, —C(O)OR⁵, —COR⁵, —SH,—SR⁵, —OR⁶ and acyl;

or R¹=L;

R² is selected from the group consisting of: H, halogen, alkyl, alkenyl,alkynyl, haloalkyl, haloalkenyl, heteroalkyl, cycloalkyl, cycloalkenyl,heterocycloalkyl, heterocycloalkenyl, aryl, heteroaryl, cycloalkylalkyl,heterocycloalkylalkyl, arylalkyl, heteroarylalkyl, arylalkenyl,cycloalkylheteroalkyl, heterocycloalkylheteroalkyl,heteroarylheteroalkyl, arylheteroalkyl hydroxy, hydroxyalkyl, alkoxy,alkoxyalkyl, alkoxyaryl, alkenyloxy, alkynyloxy, cycloalkylkoxy,heterocycloalkyloxy, aryloxy, heteroaryloxy, arylalkyloxy, amino,alkylamino, aminoalkyl, acylamino, arylamino, phenoxy, benzyloxy, COOH,alkoxycarbonyl, alkylaminocarbonyl, sulfonyl, alkylsulfonyl,alkylsulfinyl, arylsulfonyl, arylsulfinyl, aminosulfonyl, SR⁶ and acyl,each of which may be unsubstituted or substituted with one or moresubstituents independently selected from the group consisting of:halogen, ═O, ═S, —CN, —NO₂, —CF₃, —OCF₃, alkyl, alkenyl, alkynyl,haloalkyl, haloalkenyl, haloalkynyl, heteroalkyl, cycloalkyl,cycloalkenyl, heterocycloalkyl, heterocycloalkenyl, aryl, heteroaryl,hydroxy, hydroxyalkyl, alkoxy, alkoxyalkyl, alkoxyaryl,alkoxyheteroaryl, alkenyloxy, alkynyloxy, cycloalkyloxy,cycloalkenyloxy, heterocycloalkyloxy, heterocycloalkenyloxy, aryloxy,heteroaryloxy, arylalkyl, heteroarylalkyl, arylalkyloxy, amino,alkylamino, acylamino, aminoalkyl, arylamino, sulfonyl, alkylsulfonyl,arylsulfonyl, aminosulfonyl, aminoalkyl, alkoxyalky, —COOH, —COR⁵,—C(O)OR⁵, —SH, —SR⁶, —OR⁶ and acyl;

or R²=L;

R³ is selected from the group consisting of H, C₁-C₆ alkyl, and acyl; ora metal ion selected from sodium, calcium, magnesium;

X and Y are the same or different and are independently selected fromthe group consisting of: H, halogen, —CN, —NO₂, —CF₃, —OCF₃, alkyl,alkenyl, alkynyl, haloalkyl, haloalkenyl, haloalkynyl, heteroalkyl,cycloalkyl, cycloalkenyl, heterocycloalkyl, heterocycloalkenyl, aryl,heteroaryl, hydroxy, hydroxyalkyl, alkoxy, alkoxyalkyl, alkoxyaryl,alkoxyheteroaryl, alkenyloxy, alkynyloxy, cycloalkyloxy,cycloalkenyloxy, heterocycloalkyloxy, heterocycloalkenyloxy, aryloxy,heteroaryloxy, arylalkyl, heteroarylalkyl, arylalkyloxy, amino,alkylamino, acylamino, aminoalkyl, arylamino, sulfonyl, alkylsulfonyl,arylsulfonyl, aminosulfonyl, aminoalkyl, alkoxyalky, —COOH, —C(O)OR⁵,—COR⁵, —SH, —SR⁶, acyl and —NR⁷R⁸;

R⁴ is selected from the group consisting of: H, alkyl, alkenyl, alkynyl,haloalkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl,cycloalkylalkyl, heterocycloalkylalkyl, arylalkyl, heteroarylalkyl andacyl;

Each R⁵ is independently selected from the group consisting of: alkyl,alkenyl, alkynyl, haloalkyl, heteroalkyl, cycloalkyl, heterocycloalkyl,aryl, heteroaryl, cycloalkylalkyl, heterocycloalkylalkyl, arylalkyl,heteroarylalkyl and acyl;

Each R⁶ is independently selected from the group consisting of: alkyl,alkenyl, alkynyl, haloalkyl, heteroalkyl, cycloalkyl, heterocycloalkyl,aryl, heteroaryl, cycloalkylalkyl, heterocycloalkylalkyl, arylalkyl,heteroarylalkyl and acyl;

Each R⁷ and R⁸ are each independently selected from the group consistingof: H, alkyl, alkenyl, alkynyl, haloalkyl, heteroalkyl, cycloalkyl,heterocycloalkyl, aryl, heteroaryl, cycloalkylalkyl,heterocycloalkylalkyl, arylalkyl, heteroarylalkyl and acyl;

L is selected from the group consisting of:

-   -   a) L=Cy-L¹-W—    -   Wherein        -   Cy is C₁-C₁₅ alkyl, aminoalkyl, heterocycloalkyl,            cycloalkyl, aryl, aryloxy or heteroaryl any of which may be            optionally substituted one or more substituents            independently selected from the group consisting of:            halogen, ═O, ═S, —CN, —NO₂, —CF₃, —OCF₃, alkyl, alkenyl,            alkynyl, haloalkyl, haloalkenyl, haloalkynyl, heteroalkyl,            cycloalkyl, cycloalkenyl, heterocycloalkyl,            heterocycloalkenyl, aryl, heteroaryl, hydroxy, hydroxyalkyl,            alkoxy, alkoxyalkyl, alkoxyaryl, alkoxyheteroaryl,            alkenyloxy, alkynyloxy, cycloalkyloxy, cycloalkenyloxy,            heterocycloalkyloxy, heterocycloalkenyloxy, aryloxy,            heteroaryloxy, arylalkyl, heteroarylalkyl, arylalkyloxy,            amino, alkylamino, acylamino, aminoalkyl, arylamino,            sulfonyl, alkylsulfonyl, arylsulfonyl, aminosulfonyl,            aminoalkyl, alkoxyalky, —COOH, —C(O)OR⁵, —COR⁵, —SH, —SR⁶,            —OR⁶ and acyl.        -   L¹ is selected from the group consisting of C₁-C₅ alkyl,            which may be optionally substituted with one or more            substituents independently selected from the group            consisting of: halogen; ═O; ═S; —CN; —NO₂; alkyl, alkoxy,            acylamino, and alkylamino;        -   W is selected from the group consisting of a single bond,            —O—, —S—, —S(O)—, —S(O)₂—, —N(R⁹)—, —C(O)N(R⁹)—, —SO₂N(R⁹)—,            N(R⁹)C(O)—, N(R⁹)SO₂—, and —N(R⁹)—C(O)—N(R¹⁰)—;    -   b) L=Cy-L¹-W-L²    -   Wherein,        -   Cy is C₁-C₁₅ alkyl, aminoalkyl, heterocycloalkyl,            cycloalkyl, aryl, aryloxy or heteroaryl any of which may be            optionally substituted one or more substituents            independently selected from the group consisting of:            halogen, ═O, ═S, —CN, —NO₂, —CF₃, —OCF₃, alkyl, alkenyl,            alkynyl, haloalkyl, haloalkenyl, haloalkynyl, heteroalkyl,            cycloalkyl, cycloalkenyl, heterocycloalkyl,            heterocycloalkenyl, aryl, heteroaryl, hydroxy, hydroxyalkyl,            alkoxy, alkoxyalkyl, alkoxyaryl, alkoxyheteroaryl,            alkenyloxy, alkynyloxy, cycloalkyloxy, cycloalkenyloxy,            heterocycloalkyloxy, heterocycloalkenyloxy, aryloxy,            heteroaryloxy, arylalkyl, heteroarylalkyl, arylalkyloxy,            amino, alkylamino, acylamino, aminoalkyl, arylamino,            sulfonyl, alkylsulfonyl, arylsulfonyl, aminosulfonyl,            aminoalkyl, alkoxyalky, —COOH, C(O)OR⁶, —COR⁵, —SH, —SR⁶,            —OR⁶ and acyl;        -   L¹ and L² are the same or different and independently C₁-C₅            alkyl, which may be optionally substituted with one or more            substituents independently selected from the group            consisting of: halogen; ═O; ═S; —CN; —NO₂; —CF₃, —OCF₃,            alkyl, alkoxy, acylamino and alkylamino;        -   W is selected from the group consisting of a single bond,            —O—, —S—, —S(O)—, —S(O)₂—, —N(R⁹)—, —C(O)N(R⁹)—, —SO₂N(R⁹)—,            N(R⁹)C(O)—, N(R⁹)SO₂—, and —N(R⁹)—C(O)—N(R¹⁰)—;    -   c) L=Cy-(CH₂)m-W—    -   Wherein,        -   Cy is C₁-C₁₅ alkyl, aminoalkyl, heterocycloalkyl,            cycloalkyl, aryl, aryloxy or heteroaryl, any of which may be            optionally substituted one or more substituents            independently selected from the group consisting of:            halogen, ═O, ═S, —CN, —NO₂, —CF₃, —OCF₃, alkyl, alkenyl,            alkynyl, haloalkyl, haloalkenyl, haloalkynyl, heteroalkyl,            cycloalkyl, cycloalkenyl, heterocycloalkyl,            heterocycloalkenyl, aryl, heteroaryl, hydroxy, hydroxyalkyl,            alkoxy, alkoxyalkyl, alkoxyaryl, alkoxyheteroaryl,            alkenyloxy, alkynyloxy, cycloalkyloxy, cycloalkenyloxy,            heterocycloalkyloxy, heterocycloalkenyloxy, aryloxy,            heteroaryloxy, arylalkyl, heteroarylalkyl, arylalkyloxy,            amino, alkylamino, acylamino, aminoalkyl, arylamino,            sulfonyl, alkylsulfonyl, arylsulfonyl, aminosulfonyl,            aminoalkyl, alkoxyalky, —COOH, C(O)OR⁶, —COR⁵, —SH, —SR⁶,            —OR⁶ and acyl;        -   m is 0, 1, 2, 3, 4 or 5;        -   W is selected from the group consisting of a single bond,            —O—, —S—, —S(O)—, —S(O)₂—, —N(R⁹)—, —C(O)N(R⁹)—, —SO₂N(R⁹)—,            N(R⁹)C(O)—, N(R⁹)SO₂—, and —N(R⁹)—C(O)—N(R¹⁰)—;    -   d) L=L¹-W-L²        -   L¹ and L² are the same or different and independently            selected from C₁-C₅ alkyl, which may be optionally            substituted one or more substituents independently selected            from the group consisting of: halogen; ═O; ═S; —CN; —NO₂;            —CF₃, —OCF₃, alkyl, alkoxy, acylamino, alkylamino;        -   W is selected from the group consisting of a single bond,            —O—, —S—, —S(O)—, —S(O)₂—, —N(R⁹)—, —C(O)N(R⁹)—, —SO₂N(R⁹)—,            N(R⁹)C(O)—, N(R⁹)SO₂—, and —N(R⁹)—C(O)—N(R¹⁰)—;

R⁹ and R¹⁰ are the same or different and are independently selected fromH, C₁-C₆ alkyl, C₄-C₉ cycloalkyl, C₄-C₉ heterocycloalkyl, aryl,heteroaryl, arylalkyl, heteroarylalkyl; and acyl;

Z is a single bond or is selected from —CH₂—, —CH₂CH₂—, —CH═CH—, C₃-C₆cycloalkyl, unsubstituted or substituted with one or more substituentsindependently selected from the group consisting of C₁-C₄ alkyl;

or a pharmaceutically acceptable salt thereof.

As used herein, the term unsubstituted means that there is nosubstituent or that the only substituents are hydrogen.

“Halogen” represents chlorine, fluorine, bromine or iodine.

“Alkyl” as a group or part of a group refers to a straight or branchedaliphatic hydrocarbon group, preferably a C₁-C₁₄ alkyl, more preferablyC₁-C₁₀ alkyl, most preferably C₁-C₆ unless otherwise noted. Examples ofsuitable straight and branched C₁-C₆ alkyl substituents include methyl,ethyl, n-propyl, 2-propyl, n-butyl, sec-butyl, t-butyl, hexyl, and thelike.

“Alkylamino” includes both monoalkylamino and dialkylamino, unlessspecified. “Monoalkylamino” means a —NH-Alkyl group, “Dialkylamino”means a —N(alkyl)₂ group, in which the alkyl is as defined as above. Thealkyl group is preferably a C₁-C₆ alkyl group.

“Arylamino” includes both mono-arylamino and di-arylamino unlessspecified. Mono-arylamino means a group of formula aryl NH—,di-arylamino means a group of formula (aryl₂)N— where aryl is as definedherein.

“Acyl” means an alkyl-CO— group in which the alkyl group is as describedherein. Examples of acyl include acetyl and benzoyl. The alkyl group ispreferably a C₁-C₆ alkyl group.

“Alkenyl” as group or part of a group denotes an aliphatic hydrocarbongroup containing at least one carbon-carbon double bond and which may bestraight or branched preferably having 2-14 carbon atoms, morepreferably 2-12 carbon atoms, most preferably 2-6 carbon atoms, in thechain. The group may contain a plurality of double bonds in the normalchain and the orientation about each is independently E or Z. Exemplaryalkenyl group include, but are not limited to, ethenyl and propenyl.

“Alkoxy” refers to an —O-alkyl group in which alkyl is defined herein.Preferably the alkoxy is a C₁-C₆alkoxy. Examples include, but are notlimited to, methoxy and ethoxy.

“Alkenyloxy” refers to an —O— alkenyl group in which alkenyl is asdefined herein. Preferred alkenyloxy groups are C₁-C₆ alkenyloxy groups.

“Alkynyloxy” refers to an —O-alkynyl group in which alkynyl is asdefined herein, Preferred alkynyloxy groups are C₁-C₆ alkynyloxy groups.

“Alkoxycarbonyl” refers to an —C(O)—O-alkyl group in which alkyl is asdefined herein. The alkyl group is preferably a C₁-C₆ alkyl group.Examples include, but not limited to, methoxycarbonyl andethoxycarbonyl.

“Akylsulfinyl” means a —S(O)-alkyl group in which alkyl is as definedabove. The alkyl group is preferably a C₁-C₆ alkyl group. Exemplaryalkylsulfinyl groups include, but not limited to, methylsulfinyl andethylsulfinyl.

“Alkylsulfonyl” refers to a —S(O)₂-alkyl group in which alkyl is asdefined above. The alkyl group is preferably a C₁-C₈ alkyl group.Examples include, but not limited to methylsulfonyl and ethylsulfonyl.

“Alkynyl” as a group or part of a group means an aliphatic hydrocarbongroup containing a carbon-carbon trip bond and which may be straight orbranched preferably having from 2-14 carbon atoms, more preferably 2-12carbon atoms in the chain, preferably 2-6 carbon atoms in the chain.Exemplary structures include, but not limited to, ethynyl and propynyl.

“Alkylaminocarbonyl” refers to an alkylamino-carbonyl group in whichalkylamino is as defined above.

“Cycloalkyl” refers to a saturated or partially saturated, monocyclic orfused or spiro polycyclic, carbocycle preferably containing from 3 to 9carbons per ring, such as cyclopropyl, cyclobutyl, cyclopentyl,cyclohexyl and the like, unless otherwise specified.

The above discussion of alkyl and cycloalkyl substituents also appliesto the alkyl portions of other substituents, such as without limitation,alkoxy, alkyl amines, alkyl ketones, arylalkyl, heteroarylalkyl,alkylsulfonyl and alkyl ester substituents and the like.

“Cycloalkylalkyl” means a cycloalkyl-alkyl- group in which thecycloalkyl and alkyl moieties are as previously described. Exemplarymonocycloalkylalkyl groups include cyclopropylmethyl, cyclopentylmethyi,cyciohexylmethyl and cylcoheptylmethyl.

“Heterocycloalkyl” refers to an ring containing from at least oneheteroatom selected from nitrogen, sulfur, oxygen, preferably from 1 to3 heteroatoms. Each ring is preferably from 3 to 4 membered, morepreferably 4 to 7 membered. Examples of suitable heterocycloalkylsubstituents include pyrrolidyl, tetrahydrofuryl, tetrahydrothiofuranyl,piperidyl, piperazyl, tetrahydropyranyl, morphilino, 1,3-diazapane,1,4-diazapane, 1,4-oxazepane, and 1,4-oxathiapane.

“Heterocycloalkenyl” refers to a heterocycloalkyl as described above butcontaining at least one double bond.

“Heterocycloalkylalkyl” refers to a heterocycloalkyl-alkyl group inwhich the heterocycloalkyl and alkyl moieties are as previouslydescribed. Exemplary heterocycloalkylalkyl groups include(2-tetrahydrofuryl)methyl, (2-tetrahydrothiofuranyl)methyl.

“Heteroalkyl” refers to a straight- or branched-chain alkyl grouppreferably having from 2 to 14 carbons, more preferably 2 to 10 atoms inthe chain, one or more of which is a heteroatom selected from S, O, andN. Exemplary heteroalkyls include alkyl ethers, secondary and tertiaryalkyl amines, alkyl sulfides, and the like.

“Aryl” as a group or part of a group denotes (i) an optionallysubstituted monocyclic, or fused polycyclic, aromatic carbocycle (ringstructure having ring atoms that are all carbon) preferably having from5 to 12 atoms per ring. Examples of aryl groups include phenyl,naphthyl, and the like; (ii) an optionally substituted partiallysaturated bicyclic aromatic carbocyclic moiety in which a phenyl and aC₅₋₇ cycloalkyl or C₅₋₇ cycloalkenyl group are fused together to form acyclic structure, such as tetrahydronaphthyl, indenyl or indanyl. Thearyl group may be substituted by one or more substituent groups.

“Arylalkenyl” means an aryl-alkenyl- group in which the aryl and alkenylare as previously described. Exemplary arylalkenyl groups includephenylallyl.

“Arylalkyl” means an aryl-alkyl- group in which the aryl and alkylmoieties are as previously described. Preferred arylalkyl groupscontains a C₁₋₅ alkyl moiety. Exemplary arylalkyl groups include benzyl,phenethyl and naphthelenemethyl.

“Cycloalkenyl” means an optionally substituted non-aromatic monocyclicor multicyclic ring system containing at least one carbon-carbon doublebond and preferably having from 5-10 carbon atoms per ring. Exemplarymonocyclic cycloalkenyl rings include cyclopentenyl, cyclohexenyl orcycloheptenyl. The cycloalkenyl group may be substituted by one or moresubstituent groups.

“Heteroaryl” refers to a monocyclic, or fused polycyclic, aromaticheterocycle (ring structure preferably having a 5 to 7 member aromaticring containing one or more heteroatoms selected from N, O and S).Typical heteroaryl substituents include furyl, thienyl, pyrrole,pyrazole, triazole, thiazole, oxazole, pyridine, pyrimidine, isoxazolyl,pyrazine, indole, benzimidazole, and the like.

“Heteroarylalkyl” means a heteroaryl-alkyl group in which the heteroaryland alkyl moieties are as previously described. Preferredheteroarylalkyl groups contain a lower alkyl moiety. Exemplaryheteroarylalkyl groups include pyridylmethyl.

“Lower alkyl” as a group means unless otherwise specified, an aliphatichydrocarbon group which may be straight or branched having 1 to 6 carbonatoms in the chain, more preferably 1 to 4 carbons such as methyl,ethyl, propyl (n-propyl or isopropyl) or butyl (n-butyl, isobutyl ortertiary-butyl).

In Formula I, as well as in Formulae Ia-Ib defining sub-sets ofcompounds within Formula I, there is shown a benzimidazole ring system.Within this ring system, there are substitutable positions at the 4-,5-, 6-, and 7-ring positions. In each of Formulae I, la, and Ib, thereis a requirement for attachment of an acidic moiety at one of the ringpositions. This acidic moiety may be provided by but is not limited togroups containing, a hydroxamic acid or salt derivatives of such acidwhich when hydrolyzed would provide the acidic moiety. In someembodiments the acidic moiety may be attached to the ring positionthrough an alkylene group such as —CH₂— or —CH₂CH₂—, or an alkenyl groupsuch as —CH═CH—. Preferred positions for attachment of the acidic moietyare the 5- and 6-ring positions.

It is understood that included in the family of compounds of Formula Iare isomeric forms including diastereoisomers, enantiomers, tautomers,and geometrical isomers in “E” or “Z” configurational isomer or amixture of E and Z isomers. It is also understood that some isomericforms such as diastereomers, enantiomers, and geometrical isomers can beseparated by physical and/or chemical methods and by those skilled inthe art.

Some of the compounds of the disclosed embodiments may exist as singlestereoisomers, racemates, and/or mixtures of enantiomers and/ordiastereomers. All such single stereoisomers, racemates and mixturesthereof are intended to be within the scope of the subject matterdescribed and claimed.

Additionally, Formula I is intended to cover, where applicable, solvatedas well as unsolvated forms of the compounds. Thus, each formulaincludes compounds having the indicated structure, including thehydrated as well as the non-hydrated forms.

In addition to compounds of the Formula I, the HDAC inhibiting agents ofthe various embodiments include pharmaceutically acceptable salts,prodrugs, and active metabolites of such compounds, and pharmaceuticallyacceptable salts of such metabolites.

The term “Pharmaceutically acceptable salts” refers to salts that retainthe desired biological activity of the above-identified compounds, andinclude pharmaceutically acceptable acid addition salts and baseaddition salts. Suitable pharmaceutically acceptable acid addition saltsof compounds of Formula I may be prepared from an inorganic acid or froman organic acid. Examples of such inorganic acids are hydrochloric,sulfuric, and phosphoric acid. Appropriate organic acids may be selectedfrom aliphatic, cycloaliphatic, aromatic, heterocyclic carboxylic andsulfonic classes of organic acids, examples of which are formic, acetic,propionic, succinic, glycolic, gluconic, lactic, malic, tartaric,citric, fumaric, maleic, alkyl sulfonic, arylsulfonic. Suitablepharmaceutically acceptable base addition salts of compounds of FormulaI include metallic salts made from lithium, sodium, potassium,magnesium, calcium, aluminium, and zinc, and organic salts made fromorganic bases such as choline, diethanolamine, morpholine. Otherexamples of organic salts are: ammonium salts, quaternary salts such astetramethylammonium salt; amino acid addition salts such as salts withglycine and arginine. Additional information on pharmaceuticallyacceptable salts can be found in Remington's Pharmaceutical Sciences,18th Edition, Mack Publishing Co., Easton, Pa. 1990. In the case ofagents that are solids, it is understood by those skilled in the artthat the inventive compounds, agents and salts may exist in differentcrystalline or polymorphic forms, all of which are intended to be withinthe scope of the present invention and specified formulae.

“Prodrug” means a compound which is convertible in vivo by metabolicmeans (e.g. by hydrolysis, reduction or oxidation) to a compound offormula I. For example an ester prodrug of a compound of formula Icontaining a hydroxyl group may be convertible by hydrolysis in vivo tothe parent molecule. Suitable esters of compounds of formula (I)containing a hydroxyl group, are for example acetates, citrates,lactates, tartrates, malonates, oxalates, salicylates, propionates,succinates, fumarates, maleates, methylene-bis-β-hydroxynaphthoates,gestisates, isethionates, di-p-toluoyltartrates, methanesulphonates,ethanesulphonates, benzenesulphonates, p-toluenesulphonates,cyclohexylsulphamates and quinates. As another example an ester prodrugof a compound of formula I containing a carboxy group may be convertibleby hydrolysis in vivo to the parent molecule. (Examples of esterprodrugs are those described by F. J. Leinweber, Drug Metab. Res.,18:379, 1987).

Possible HDAC inhibiting agents include those having an IC50 value of 1μM or less.

Administration of compounds within Formula I to humans can be by any ofthe accepted modes for enteral administration such as oral or rectal, orby parenteral administration such as subcutaneous, intramuscular,intravenous and intradermal routes. Injection can be bolus or viaconstant or intermittent infusion. The active compound is typicallyincluded in a pharmaceutically acceptable carrier or diluent and in anamount sufficient to deliver to the patient a therapeutically effectivedose. In various embodiments the inhibitor compound may be selectivelytoxic or more toxic to rapidly proliferating cells, e.g. canceroustumors, than to normal cells.

The term “therapeutically effective amount” or “therapeutic amount” isan amount sufficient to effect beneficial or desired clinical results.An effective amount can be administered in one or more administrations.An effective amount is typically sufficient to palliate, ameliorate,stabilize, reverse, slow or delay the progression of the disease state.

In using the compounds of the invention they can be administered in anyform or mode which makes the compound bioavailable. One skilled in theart of preparing formulations can readily select the proper form andmode of administration depending upon the particular characteristics ofthe compound selected, the condition to be treated, the stage of thecondition to be treated and other relevant circumstances. We refer thereader to Remingtons Pharmaceutical Sciences, 18^(th) edition, MachPublishing Co. (1990) for further information.

The compounds of the present invention can be administered alone or inthe form of a pharmaceutical composition in combination with apharmaceutically acceptable carrier, diluent or excipient. The compoundsof the invention, while effective themselves, are typically formulatedand administered in the form of their pharmaceutically acceptable saltsas these forms are typically more stable, more easily crystallised andhave increased solubility.

The compounds are, however, typically used in the form of pharmaceuticalcompositions which are formulated depending on the desired mode ofadministration. As such in a further embodiment the present inventionprovides a pharmaceutical composition including a compound of Formula(I) and a pharmaceutically acceptable carrier, diluent or excipient. Thecompositions are prepared in manners well known in the art.

The compounds of the invention may be used or administered incombination with one or more additional drug(s) that arechemotherapeutic drugs or HDAC inhibitor drugs and/or procedures (e.g.surgery, radiotherapy) for the treatment of the disorder/diseasesmentioned. The components can be administered in the same formulation orin separate formulations. If administered in separate formulations thecompounds of the invention may be administered sequentially orsimultaneously with the other drug (s).

Pharmaceutical compositions of this invention for parenteral injectioncomprise pharmaceutically acceptable sterile aqueous or nonaqueoussolutions, dispersions, suspensions or emulsions as well as sterilepowders for reconstitution into sterile injectable solutions ordispersions just prior to use. Examples of suitable aqueous andnonaqueous carriers, diluents, solvents or vehicles include water,ethanol, polyols (such as glycerol, propylene glycol, polyethyleneglycol, and the like), and suitable mixtures thereof, vegetable oils(such as olive oil), and injectable organic esters such as ethyl oleate.Proper fluidity can be maintained, for example, by the use of coatingmaterials such as lecithin, by the maintenance of the required particlesize in the case of dispersions, and by the use of surfactants.

These compositions may also contain adjuvants such as preservative,wetting agents, emulsifying agents, and dispersing agents. Prevention ofthe action of microorganisms may be ensured by the inclusion of variousantibacterial and antifungal agents, for example, paraben,chlorobutanol, phenol sorbic acid, and the like. It may also bedesirable to include isotonic agents such as sugars, sodium chloride,and the like. Prolonged absorption of the injectable pharmaceutical formmay be brought about by the inclusion of agents that delay absorptionsuch as aluminium monostearate and gelatin.

If desired, and for more effective distribution, the compounds can beincorporated into slow release or targeted delivery systems such aspolymer matrices, liposomes, and microspheres.

The injectable formulations can be sterilized, for example, byfiltration through a bacterial-retaining filter, or by incorporatingsterilizing agents in the form of sterile solid compositions that can bedissolved or dispersed in sterile water or other sterile injectablemedium just prior to use.

Solid dosage forms for oral administration include capsules, tablets,pills, powders, and granules. In such solid dosage forms, the activecompound is mixed with at least one inert, pharmaceutically acceptableexcipient or carrier such as sodium citrate or dicalcium phosphateand/or a) fillers or extenders such as starches, lactose, sucrose,glucose, mannitol, and silicic acid, b) binders such as, for example,carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone,sucrose, and acacia, c) humectants such as glycerol, d) disintegratingagents such as agar-agar, calcium carbonate, potato or tapioca starch,alginic acid, certain silicates, and sodium carbonate, e) solutionretarding agents such as paraffin, f) absorption accelerators such asquaternary ammonium compounds, g) wetting agents such as, for example,cetyl alcohol and glycerol monostearate, h) absorbents such as kaolinand bentonite clay, and i) lubricants such as talc, calcium stearate,magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate,and mixtures thereof. In the case of capsules, tablets and pills, thedosage form may also comprise buffering agents.

Solid compositions of a similar type may also be employed as fillers insoft and hard-filled gelatin capsules using such excipients as lactoseor milk sugar as well as high molecular weight polyethylene glycols andthe like.

The solid dosage forms of tablets, dragees, capsules, pills, andgranules can be prepared with coatings and shells such as entericcoatings and other coatings well known in the pharmaceutical formulatingart. They may optionally contain opacifying agents and can also be of acomposition that they release the active ingredient(s) only, orpreferentially, in a certain part of the intestinal tract, optionally,in a delayed manner. Examples of embedding compositions which can beused include polymeric substances and waxes.

If desired, and for more effective distribution, the compounds can beincorporated into slow release or targeted delivery systems such aspolymer matrices, liposomes, and microspheres.

The active compounds can also be in microencapsulated form, ifappropriate, with one or more of the above-mentioned excipients.

Liquid dosage forms for oral administration include pharmaceuticallyacceptable emulsions, solutions, suspensions, syrups and elixirs. Inaddition to the active compounds, the liquid dosage forms may containinert diluents commonly used in the art such as, for example, water orother solvents, solubilizing agents and emulsifiers such as ethylalcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzylalcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol,dimethyl formamide, oils (in particular, cottonseed, groundnut, corn,germ, olive, castor, and sesame oils), glycerol, tetrahydrofurfurylalcohol, polyethylene glycols and fatty acid esters of sorbitan, andmixtures thereof.

Besides inert diluents, the oral compositions can also include adjuvantssuch as wetting agents, emulsifying and suspending agents, sweetening,flavoring, and perfuming agents.

Suspensions, in addition to the active compounds, may contain suspendingagents as, for example, ethoxylated isostearyl alcohols, polyoxyethylenesorbitol and sorbitan esters, microcrystalline cellulose, aluminiummetahydroxide, bentonite, agar-agar, and tragacanth, and mixturesthereof.

Compositions for rectal or vaginal administration are preferablysuppositories which can be prepared by mixing the compounds of thisinvention with suitable non-irritating excipients or carriers such ascocoa butter, polyethylene glycol or a suppository wax which are solidat room temperature but liquid at body temperature and therefore melt inthe rectum or vaginal cavity and release the active compound.

Dosage forms for topical administration of a compound of this inventioninclude powders, patches, sprays, ointments and inhalants. The activecompound is mixed under sterile conditions with a pharmaceuticallyacceptable carrier and any needed preservatives, buffers, or propellantswhich may be required.

A preferred dosage will be a range from about 0.01 to 300 mg perkilogram of body weight per day. A more preferred dosage will be in therange from 0.1 to 100 mg per kilogram of body weight per day, morepreferably from 0.2 to 80 mg per kilogram of body weight per day, evenmore preferably 0.2 to 50 mg per kilogram of body weight per day. Asuitable dose can be administered in multiple sub-doses per day.

As discussed above, the compounds of the embodiments disclosed inhibithistone deacetylases. The enzymatic activity of a histone deacetylasecan be measured using known methodologies [Yoshida M. et al, J. Biol.Chem., 265, 17174 (1990), J. Taunton et al, Science 1996 272: 408]. Incertain embodiments, the histone deacetylase inhibitor interacts withand reduces the activity of more than one known histone deacetylase inthe cell. In some other embodiments, the histone deacetylase inhibitorinteracts and reduces the activity of predominantly one histonedeacetylase, for example HDAC-1, HDAC-3 or HDAC-8 which belongs to ClassI HDAC enzymes [De Ruijter A. J. M. et al, Biochem. J., 370, 737-749(2003)]. Certain preferred histone deacetylase inhibitors are those thatinteract with, and reduce the activity of a histone deacetylase which isinvolved in tumorigenesis, and these compounds may be useful fortreating proliferative diseases. Examples of such cell proliferativediseases or conditions include cancer and/or any metastases, psoriasis,and restenosis. The inventive compounds may be particularly useful fortreating tumors such as breast cancer, lung cancer, ovarian cancer,prostate cancer, head and/or neck cancer, or renal, gastric, and braincancer. In addition, the inventive compounds may be useful for treatinga proliferative disease that is refractory to the treatment with otherchemotherapeutics; and for treating hyperproliferative condition such asleukemias, psoriasis, restenosis.

Additionally compounds of the various embodiments disclosed herein maybe useful for treating neurodegenerative diseases, and inflammatorydiseases and/or immune system disorders.

The disorder is preferably selected from the group consisting of cancer,inflammatory diseases and/or immune system disorders (e.g. rheumatoidarthritis, systemic lupus erythematosus), angiofibroma, cardiovasculardiseases, fibrotic diseases, diabetes, autoimmune diseases, chronic andacute neurodegenerative disease like Huntington's disease, Parkinson'sdisease, disruptions of nerval tissue and infectious diseases likefungal, bacterial and viral infections. In another embodiment thedisorder is a proliferative disorder.

The histone deacetylase inhibitors of the invention have significantantiproliferative effects and promote differentiation, cell cycle arrestin the G1 or G2 phase, and apoptosis.

Synthesis of Deacetylase Inhibitors

The agents of the various embodiments may be prepared using the reactionroutes and synthesis schemes as described below, employing thetechniques available in the art using starting materials that arereadily available. The preparation of particular compounds of theembodiments is described in detail in the following examples, but theartisan will recognize that the chemical reactions described may bereadily adapted to prepare a number of other agents of the variousembodiments. For example, the synthesis of non-exemplified compounds maybe successfully performed by modifications apparent to those skilled inthe art, e.g. by appropriately protecting interfering groups, bychanging to other suitable reagents known in the art, or by makingroutine modifications of reaction conditions. A list of suitableprotecting groups in organic synthesis can be found in T. W. Greene'sProtective Groups in Organic Synthesis, John Wiley & Sons, 1981.Alternatively, other reactions disclosed herein or known in the art willbe recognized as having applicability for preparing other compounds ofthe various embodiments.

Reagents useful for synthesizing compounds may be obtained or preparedaccording to techniques known in the art.

In the examples described below, unless otherwise indicated, alltemperatures in the following description are in degrees Celsius and allparts and percentages are by weight, unless indicated otherwise.

Various starting materials and other reagents were purchased fromcommercial suppliers, such as Aldrich Chemical Company or LancasterSynthesis Ltd., and used without further purification, unless otherwiseindicated. Tetrahydrofuran (THF) and N,N-dimethylformamide (DMF) werepurchased from Aldrich in SureSeal bottles and used as received. Allsolvents were purified by using standard methods in the art, unlessotherwise indicated.

The reactions set forth below were performed under a positive pressureof nitrogen, argon or with a drying tube, at ambient temperature (unlessotherwise stated), in anhydrous solvents, and the reaction flasks arefitted with rubber septa for the introduction of substrates and reagentsvia syringe. Glassware was oven-dried and/or heat-dried. Analyticalthin-layer chromatography was performed on glass-backed silica gel 60 F254 plates (E Merck (0.25 mm)) and eluted with the appropriate solventratios (v/v). The reactions were assayed by TLC and terminated as judgedby the consumption of starting material.

The TLC plates were visualized by UV absorption or with a p-anisaldehydespray reagent or a phosphomolybdic acid reagent (Aldrich Chemical, 20 wt% in ethanol) which was activated with heat, or by staining in iodinechamber. Work-ups were typically done by doubling the reaction volumewith the reaction solvent or extraction solvent and then washing withthe indicated aqueous solutions using 25% by volume of the extractionvolume (unless otherwise indicated). Product solutions were dried overanhydrous sodium sulfate prior to filtration, and evaporation of thesolvents was under reduced pressure on a rotary evaporator and noted assolvents removed in vacuo. Flash column chromatography [Still et al, J.Org. Chem., 43, 2923 (1978)] was conducted using E Merck-grade flashsilica gel (47-61 mm) and a silica gel:crude material ratio of about20:1 to 50:1, unless otherwise stated. Hydrogenolysis was done at thepressure indicated or at ambient pressure.

1H NMR spectra was recorded on a Bruker instrument operating at 400 MHz,and ¹³C-NMR spectra was recorded operating at 100 MHz. NMR spectra areobtained as CDCl₃ solutions (reported in ppm), using chloroform as thereference standard (7.25 ppm and 77.00 ppm) or CD₃OD (3.4 and 4.8 ppmand 49.3 ppm), or an internal tetramethylsilane standard (0.00 ppm) whenappropriate. Other NMR solvents were used as needed. When peakmultiplicities are reported, the following abbreviations are used:s=singlet, d=doublet, t=triplet, m=multiplet, br=broadened, dd=doubletof doublets, dt=doublet of triplets. Coupling constants, when given, arereported in Hertz.

Mass spectra were obtained using LC/MS either in ESI or APCI. Allmelting points are uncorrected.

All final products had greater than 90% purity (by HPLC at wavelengthsof 220 nm and 254 nm).

The following examples are intended to illustrate the embodimentsdisclosed and are not to be construed as being limitations thereto.Additional compounds, other than those described below, may be preparedusing the following described reaction scheme or appropriate variationsor modifications thereof.

Synthesis

Scheme I illustrates the procedure used for preparing compounds offormula Ib, wherein X and Y are hydrogens, compounds (VI) of formula Iacan be prepared by analogous procedure, for example, by the choice ofappropriate starting material. For example, in the case of Z is —CH═CH—and attached to C₅-position in Formula Ib, such compound(s) can besynthesized by analogous method illustrated in Scheme I starting with asubstituted cinnamic acid (e.g. trans-3-nitro-4-chloro-cinnamic acid),appropriate amine component (R¹NH₂), aldehyde or carboxylic acidcomponent (R²CHO or R²COOH), and appropriate hydroxylamine or N-alkylhydroxylamine (NHR³OH where R³ is defined as above in Formula Ia).

Specifically, the hydroxamate compounds Formula Ib can be synthesized bythe synthetic route shown in Scheme I. The reaction oftrans-4-chloro-3-nitrocinnamic acid (I) with an amine in the present ofa base (e.g. triethylamine) in an appropriate solvent (e.g. dioxane)gave (II). Treatment of (II) in methanol under acid catalysis (e.g.sulfuric acid) resulted in esterification providing (III). The nitrogroup of (Ill) can be reduced by appropriate reducing agent (e.g. tinchloride) and the resulting phenylenediamine was cyclized with analdehyde to give (V). The hydroxamate compounds (VI) were obtained by aknown synthesis method (J. Med. Chem., 2002, 45, 753-757). Analternative method for preparation of (VI) is by coupling (IV) with anappropriate acid and then cyclized by heating with acetic acid (J. Med.Chem. 2001, 44, 1516-1529).

Scheme II illustrates another alternative procedure used for preparingcompounds of formula Ib, where X and Y are hydrogens, R²=Cy-L¹-W-L². Forexample, in the case of Z is —CH═CH— and attached to C₅-position inFormula Ib, such compound(s) (XV) can be synthesized by analogous methodillustrated in Scheme II starting with appropriate (Ill), appropriateFmoc protected amino acids, appropriate acid chlorides or aldehydes, andhydroxylamine.

More specifically, for example, the hydroxamate compounds Formula Ib,where X and Y are hydrogens, R²=Cy-L¹-W-L² and Z is attached toC₅-position, can be synthesized by the synthetic route shown in SchemeII. Appropriate intermediates (III) were reduced with Tin chloride tothe corresponding diamines (VII). The coupling reaction with appropriateFmoc protected amino acids in the presence of PyBOP gave two couplingproducts (VIII) and (IX). Without further separation, (VIII) and (IX)were subjected to cyclization under acid conditions and yielded (X). Thekey intermediate (XI) can be obtained by treating (X) with 20%piperidine. Treatment of (XI) with an appropriate acid chloride or anappropriate sulfonyl chloride gave (XII) and the target compounds (XIII)were obtained by using similar method described above.

When (XI) was reacted with an appropriate aldehyde under reductionconditions (NaBH(OAc)₃/CH₃COOH), (XIV) was obtained and can betransformed to corresponding hydroxamate derivatives (XV) by the samemethods described above.

Hydroxamate compounds of Formula I can also be prepared through solidphase synthesis. Scheme III illustrates the synthesis of hydroxamatecompounds of Formula Ib. For example, in the case of Z is —CH═CH— andattached to C₅-position, in Formula Ib, such compound(s) (VI) can besynthesized by analogous method illustrated in Scheme III starting withSASRIN resin, an appropriate hydroxylamine (e.g.,O-(2,4-dimethoxy-phenyl)-hydroxylamine), an appropriate cinnamic acid(e.g., trans-4-chloro-3-nitro-cinnamic acid), an appropriate amine andan aldehyde.

Specifically, for example, the hydroxamate compounds (VI) Formula Ib canbe synthesized by the synthetic route shown in Scheme IV. The SASRINresin was treated with O-(2,4-dimethoxy-phenyl)-hydroxylamine underreductive conditions (NaBH₃CN/CH₃COOH) in an appropriate solvent gavecorresponding compound (XVI). (XVI) was reacted withtrans-4-chloro-3-nitro-cinnamic acid in the presence of4-dimethylaminopyridine to yield (XVII). Further treatment of (XVII)with appropriate amines yielded (XVIII). (XIX) was obtained by cleavageof the corresponding resin (XVIII). Without further purification, (XIX)was transformed to the corresponding hydroxamate compounds (VI) usingthe method described above.

Scheme IV illustrates another procedure for the preparation ofhydroxamate compounds of Formula I. For example, in the case of Z is—CH₂CH₂— and attached to C₅-position in Formula Ib, such compound(s) canbe synthesized by analogous method illustrated in Scheme IV startingwith appropriate intermediates (V) through reduction and then theresulting product (XX) can be transformed into corresponding hydroxamatecompounds (XXI) of Formula Ib. Compounds (XXIII) in which Z is acyclopropylene group

and attached to C₅-position in Formula Ib, can be prepared from V bytreating with (CH₃)₃S(O)I, and the resulting cyclopropyl derivatives(XXII) was converted to corresponding hydroxamate derivatives (XXIII)according to methods described above for the preparation of hydroxamicacid.

Scheme V illustrates another synthetic procedure of hydroxamatecompounds of Formula I. For example, in the case of Z is —CH═CH— andattached to C₅-position in Formula Ib, such compound(s) can besynthesized by analogous method illustrated in Scheme V starting withappropriate intermediates (II) through reduction and then the resultingproduct (XXIV) was, without further purification, cyclized to give(XXV). (XXV) was treated with an appropriate alkyl halide (e.g., benzylbromide) in the presence of an inorganic base (e.g., sodium carbonate)in an appropriate solvent to give (XXVI). Treatment of (XXVI) withhydrogen peroxide in acetic acid led to (XXVIII). Using the same methoddescribed previously, both (XXVI) and (XXVIII) were transformed intocorresponding hydroxamate compounds (XXVII), and (XXIX), respectively.

The following preparation and examples are given to enable those skilledin the art to more clearly understand and to practice the subject matterhereof. They should not be considered as limiting the scope of thedisclosure, but merely as being illustrative and representative thereof.

Example 1 Preparation ofN-Hydroxy-3-[1-(3-hydroxy-propyl)-2-(2-phenyl-propyl)-1H-benzimidazol-5-yl]-acrylamide(1) Step 1

To a pre-stirred solution of trans-4-chloro-3-nitrocinnamic acid (1.0 g,4.4 mmol) in dioxane (10 mL) was added triethylamine (2 mL),3-amino-1-propanol (1.5 mL). The resulting solution was heated to 85° C.for 19 hours and then cooled to room temperature. The solvent wasremoved under vacuum. Water (100 mL) was added to the residue and the pHwas adjusted to 1-1.5. The precipitate was collected and washed withcold water for 2 times and dried. The product3-[3-nitro-4-(hydroxypropylamine)-phenyl]-acrylic acid was obtained asyellow solid (1.10 g, 95%). MS(m/z): 267 (MH)⁺.

Step 2

Concentrated sulfuric acid (0.5 mL) was added to the solution oftrans-4-(3-hydroxypropylamine)-3-nitrocinnamic acid, (1.10 g, 3.9 mmol)and MeOH (15 mL). The resulting solution was heated to reflux for 18hours. The reaction mixture was cooled at −10° to −15° C. for 3 hours.3-[3-nitro-4-(hydroxypropylamine)-phenyl]-acrylic acid methyl ester wascollected as crystalline yellow solid (1.06 g, 91%). MS(m/z): 281 (MH)⁺.

Step 3

To a pre-stirred solution of methyltrans-4-(3-hydroxypropylamine)-3-nitrocinnamate (280 mg, 1.0 mmol) and3-phenylbutyraldehyde (500 mg, 3.4 mmol) in glacial acetic acid (5 mL),Tin chloride was added (1.18 g, 10.0 mmol). The resulting solution washeated to 45° C. for 17 hours and then cooled to room temperature. Thesolvent was removed under vacuum. Water (20 mL) and dichloromethane (20mL) was added to the residue and stirred for 30 minutes. The organiclayer was dried (MgSO₄), filtered and concentrated to an oily residue.100 mL diethyl ether was added and stirred for 4 hours. The product3-[1-(3-Hydroxy-propyl)-2-(2-phenyl-propyl)-1H-benzimidazol-5-yl]-acrylicacid methyl ester was obtained in 34.9% yield (132.0 mg). MS(m/z): 379(MH)⁺.

Step 4

Sodium methoxide (30% in methanol) (782 mg, 4.1 mmol) was added to aprestirred solution of3-[1-(3-Hydroxy-propyl)-2-(2-phenyl-propyl)-1H-benzimidazol-5-yl]-acrylicacid methyl ester (130 mg, 0.34 mmol and hydroxylamine hydrochloride(242 mg, 3.4 mmol in MeOH (1.5 mL). The reaction mixture wascontinuously stirred for 40 minutes at room temperature and then pouredinto a solution of ice-water containing 1.0 mL concentrated hydrochloricacid. The mixture was extracted with dichloromethane. The organic layerwas dried (MgSO₄), filtered and concentrated. The desired product wasseparated by reverse phase preparative HPLC. After lyopholyzation, 7.8mg (6%) ofN-Hydroxy-3-[1-(3-hydroxy-propyl)-2-(2-phenyl-propyl)-1H-benzimidazol-5-yl]-acrylamidewas obtained as powder. HPLC: 96%; t_(R)=(LC/PDA: Phenomenex Luna C182.0×150 mm 5μ column; 0.8 mL/min, gradient 5-65% B over 15.5 min,Solvent A: H₂O with 0.1% trifluoroacetic acid; Solvent B: Acetonitrilewith 0.1% trifluoroacetic acid; UV 254): 7.22 min; 92%. ¹H NMR (400 MHz,DMSO-d₆) δ 1.35 (3H, d, J=6.5 Hz), 1.83 (2H, m), 3.00-4.00 (6H, m), 4.33(2H, t, J=7.1 Hz), 6.55 (1H, d, J=15.8 Hz), 7.19-7.33 (5H, m), 7.62 (1H,d, J=15.8 Hz), 7.70 (1H, d, J=8.60 Hz), 7.82 (1H, d, J=8.60 Hz), 7.92(1H, s), 10.15 (1H, bs), 10.33 (1H, bs). MS(m/z): 380 [MH]⁺.

Example 2 Preparation ofN-Hydroxy-3-[1-(3,4,5-trimethoxybenzyl)-2-(2-phenyl-ethyl)-1H-benzimidazol-5-yl]-acrylamide(2)

The titled compound (2) was prepared according to the proceduresdescribed in Example 1, by using appropriate starting materials. HPLC:91%; t_(R)=(LC/PDA: Phenomenex Luna C18 2.0×150 mm 5μ column; 0.8mL/min, gradient 5-65% B over 15.5 min, Solvent A: H₂O with 0.1%trifluoroacetic acid; Solvent B: Acetonitrile with 0.1% trifluoroaceticacid; UV 254): 7.22 min. ¹H NMR (400 MHz, DMSO-d₆) δ 3.08 (2H, t, J=7.72Hz), 3.48 (2H, t, 7.72 Hz), 3.63 (3H, s), 3.67 (6H, s), 5.58 (2H, s),6.59 (2H, s), 7.22-7.31 (7H, m), 7.63 (1H, d, J=15.78 Hz), 7.71 (1H, d,J=8.76 Hz), 7.83 (1H, d, J=8.76 Hz), 7.98 (1H, s), 11.00 (2H, bs).MS(m/z): 488 [MH]⁺.

Example 3 Preparation ofN-Hydroxy-3-[2-(4-benzyloxy-3-methoxy-phenyl)-1-methyl-1H-benzimidazole-5-yl]-acrylamide(3)

The titled compound (3) was prepared according to the proceduresdescribed in Example 1, by using appropriate starting materials. HPLC:92%; t_(R)=(LC/PDA: Phenomenex Luna C18 2.0×150 mm 5μ column; 0.8mL/min, gradient 5-65% B over 15.5 min, Solvent A: H₂O with 0.1%trifluoroacetic acid; Solvent B: Acetonitrile with 0.1% trifluoroaceticacid; UV 254): 7.32 min. ¹H NMR (400 MHz, DMSO-d₆) δ 3.87 (3H, s), 4.01(3H, s), 5.24 (2H, s), 6.56 (1H, d=15.80 Hz), 7.32-7.50 (8H, m), 7.74(1H, d, J=8.72 Hz), 7.88 (1H, d, J=8.72 Hz), 7.94 (1H, s), 10.85 (1H,bs). MS (m/z): 431 [MH]⁺.

Example 4 Preparation ofN-Hydroxy-3-[2-(4-benzyloxy-3-methoxy-phenyl)-1-(3-hydroxy-propyl)-1H-benzimidazole-5-yl]-acrylamide(4)

The titled compound (4) was prepared according to the proceduresdescribed in Example 1, by using appropriate starting materials. HPLC:95%; t_(R)=(LC/PDA: Phenomenex Luna C18 2.0×150 mm 5μ column; 0.8mL/min, gradient 5-65% B over 15.5 min, Solvent A: H₂O with 0.1%trifluoroacetic acid; Solvent B: Acetonitrile with 0.1% trifluoroaceticacid; UV 254): 6.82 min. ¹H NMR (400 MHz, DMSO-d₆) δ 1.96 (2H, m), 3.88(3H, s), 4.48 (2H, t, J=7.12 Hz), 5.24 (2H, s), 6.56 (1H, d, J=15.76Hz), 7.32-7.50 (8H, m), 7.65 (1H, d, J=15.76 Hz), 7.74 (1H, d, J=8.60Hz), 7.91 (1H, d, J=8.60 Hz), 7.95 (1H, s), 10.85 (1H, bs). MS (m/z):474 [MH]⁺.

Example 5 Preparation ofN-Hydroxy-3-[1-(2-hydroxy-ethyl)-2-(4-methoxy-phenyl)-1H-benzimidazole-5-yl]-acrylamide(5)

The titled compound (5) was prepared according to the proceduresdescribed in Example 1, by using appropriate starting materials. HPLC:98%; t_(R)=(LC/PDA: Phenomenex Luna C18 2.0×150 mm 5μ column; 0.8mL/min, gradient 5-65% B over 15.5 min, Solvent A: H₂O with 0.1%trifluoroacetic acid; Solvent B: Acetonitrile with 0.1% trifluoroaceticacid; UV 254): 4.12 min. ¹H NMR (400 MHz, DMSO-d₆) δ 3.80 (2H, t, J=5.36Hz), 3.87 (3H, s), 4.39 (2H, t, J=5.36 Hz), 6.56 (1H, d, 15.72 Hz), 7.17(2H, d, J=8.88 Hz), 7.61 (1H, d, J=8.52 Hz), 7.62 (1H, d, J=15.72 Hz),7.78 (1H, d, J=8.52 Hz), 7.88 (1H, d, J=8.88 Hz), 7.90 (1H, s), 10.77(1H, bs). MS (m/z): 354 [MH]⁺.

Example 6 Preparation ofN-Hydroxy-3-[1-(2,3-hydroxy-propyl)-2-(4-methoxy-phenyl)-1H-benzimidazole-5-yl]-acrylamide(6)

The titled compound (6) was prepared according to the proceduresdescribed in Example 1, by using appropriate starting materials. HPLC:98%, t_(R)=(LC/PDA: Phenomenex Luna C18 2.0×150 mm 5μ column; 0.8mL/min, gradient 5-65% B over 15.5 min, Solvent A: H₂O with 0.1%trifluoroacetic acid; Solvent B: Acetonitrile with 0.1% trifluoroaceticacid; UV 254): 3.39 min. NMR (400 MHz, DMSO-d₆) δ 3.90 (3H, s), 4.01(1H, m), 4.35 (2H, m), 4.58 (2H, dd, J=2.48 and 14.48 Hz), 6.62 (1H, d,J=15.84 Hz), 7.27 (2H, d, J=8.92 Hz), 7.68 (1H, d, J=15.84 Hz), 8.01(4H, m), 10.13 (1H, bs). MS (m/z): 383 [M]⁺.

Example 7 Preparation ofN-Hydroxy-3-[2-(4-benzyloxy-3-methoxy-phenyl)-1-(2,3-hydroxy-propyl)-1H-benzimidazole-5-yl]-acrylamide(7)

The titled compound (7) was prepared according to the proceduresdescribed in Example 1, by using appropriate starting materials. HPLC:100%, t_(R)=(LC/PDA: Phenomenex Luna C18 2.0×150 mm 5μ column; 0.8mL/min, gradient 5-65% B over 15.5 min, Solvent A: H₂O with 0.1%trifluoroacetic acid; Solvent B: Acetonitrile with 0.1% trifluoroaceticacid; UV 254): 2.06 min. NMR (400 MHz, DMSO-d₆) δ 4.04-4.38 (3H, m),4.05 (3H, s), 4.49 (2H, m), 5.22 (2H, s), 6.55 (1H, d, J=15.72 Hz),7.29-7.94 (11H, m), 8.01 (1H, s). MS (m/z): 490 [MH]⁺.

Example 8 Preparation ofN-Hydroxy-3-[1-(23-hydroxy-propyl)-2-(2-pyridyl)-1H-benzimidazol-5-yl]-acrylamide(9)

The titled compound (9) was prepared according to the proceduresdescribed in Example 1, by using appropriate starting materials. HPLC:93.7%, t_(R)=(LC/PDA: Phenomenex Luna C18 2.0×150 mm 5μ column; 0.8mL/min, gradient 5-65% B over 15.5 min, Solvent A: H₂O with 0.1%trifluoroacetic acid; Solvent B: Acetonitrile with 0.1% trifluoroaceticacid; UV 254): 2.61 min. NMR (400 MHz, DMSO-d₆) δ 3.20-3.37 (4H, m),3.90 (1H, m), 4.90-4.95 (2H, m), 6.54 (1H, d, J=15.52 Hz), 7.98 (1H, s),8.04 (1H, m), 8.27 (1H, m), 9.73 (1H, d, J=8.0 Hz). MS (m/z): 355 [MH]).

Example 9 Preparation ofN-Hydroxy-3-[1-(2-hydroxy-ethyl)-2-(4-pyridyl)-1H-benzimidazol-5-yl]-acrylamide(10)

The titled compound (10) was prepared according to the proceduresdescribed in Example 1, by using appropriate starting materials. HPLC:97.0%; t_(R)=(LC/PDA: Phenomenex Luna C18 2.0×150 mm 5μ column; 0.8mL/min, gradient 5-65% B over 15.5 min, Solvent A: H₂O with 0.1%trifluoroacetic acid; Solvent B: Acetonitrile with 0.1% trifluoroaceticacid; UV 254): 1.14 min. NMR (400 MHz, DMSO-d₆) δ 3.78 (2H, t, J=5.80Hz), 4.43 (2H, t, J=5.80 Hz), 6.50 (1H, d, J=15.80 Hz), 7.82 (2H, d,J=8.56 Hz), 7.94 (1H, s), 8.00 (2H, d, J=5.97 Hz), 8.81 (2H, d, J=5.97Hz). MS (m/z): 325 [MH]⁺.

Example 10 Preparation ofN-Hydroxy-3-[1-(3-hydroxy-propyl)-2-(4-pyridyl)-1H-benzimidazol-5-yl]-acrylamide(11)

The titled compound (11) was prepared according to the proceduresdescribed in Example 1, by using appropriate starting materials. HPLC:98.2%; t_(R)=(LC/PDA: Phenomenex Luna C18 2.0×150 mm 5μ column; 0.8mL/min, gradient 5-65% B over 15.5 min, Solvent A: H₂O with 0.1%trifluoroacetic acid; Solvent B: Acetonitrile with 0.1% trifluoroaceticacid; UV 254): 2.61 min. NMR (400 MHz, DMSO-d₆) δ 1.91 (2H, m), 3.37(2H, t, J=5.84 Hz), 4.49 (2H, t, J=7.84 Hz), 6.54 (1H, d, J=15.52 Hz),7.98 (1H, s), 8.06 (2H, d, J=6.26 Hz), 8.90 (2H, d, J=626 Hz). MS (m/z):339 [MH]⁺.

Example 11 Preparation ofN-Hydroxy-3-[1-(3-pyridylmethyl)-2-(2-phenyl-ethyl)-1H-benzimidazol-5-yl]-acrylamide(12)

The titled compound (12) was prepared according to the proceduresdescribed in Example 1, by using appropriate starting materials. HPLC:97.9%; t_(R)=(LC/PDA: Phenomenex Luna C18 2.0×150 mm 5μ column; 0.8mL/min, gradient 5-65% B over 15.5 min, Solvent A: H₂O with 0.1%trifluoroacetic acid; Solvent B: Acetonitrile with 0.1% trifluoroaceticacid; UV 254): 3.32 min. NMR (400 MHz, DMSO-d₆) δ 3.11 (2H, t, J=8.40Hz), 5.71 (2H, s), 6.51 (1H, d, J=15.80 Hz), 7.20-7.31 (6H, m), 7.43(1H, m), 7.40-7.57 (4H, m), 7.94 (1H, s), 8.57 (1H, s). MS (m/z): 399[MH]⁺.

Example 12 Preparation ofN-Hydroxy-3-[1-(3-hydroxy-propyl)-2-(2-pyridyl)-1H-benzimidazol-5-yl]-acrylamide(13)

The titled compound (13) was prepared according to the proceduresdescribed in Example 1, by using appropriate starting materials. HPLC:98.3%; t_(R)=(LC/PDA: Phenomenex Luna C18 2.0×150 mm 5μ column; 0.8mL/min, gradient 5-65% B over 15.5 min, Solvent A: H₂O with 0.1%trifluoroacetic acid; Solvent B: Acetonitrile with 0.1% trifluoroaceticacid; UV 254): 3.37 min. NMR (400 MHz, DMSO-d₆) δ 1.98 (2H, m), 3.30(2H, m), 4.86 (2H, t, J=7.00 Hz), 6.51 (1H, d, J=15.76 Hz), 7.77 (2H, d,J=8.56 Hz), 7.94 (1H, s), 8.05 (1H, m), 8.30 (1H, d, J=7.92 Hz), 8.78(1H, d, J=4.28 Hz). MS (m/z): 339 [MH]⁺.

Example 13 Preparation ofN-Hydroxy-3-[1-(3-hydroxy-propyl)-2-phenethyl-1H-benzimidazol-5-yl]-acrylamide(14)

The titled compound (14) was prepared according to the proceduresdescribed in Example 1, by using appropriate starting materials. HPLC:97.3%; t_(R)=(LC/PDA: Phenomenex Luna C18 2.0×150 mm 5μ column; 0.8mL/min, gradient 5-65% B over 15.5 min, Solvent A: H₂O with 0.1%trifluoroacetic acid; Solvent B: Acetonitrile with 0.1% trifluoroaceticacid; UV 254): 2.63 min. NMR (400 MHz, DMSO-d₆) δ 1.87 (2H, m), 3.18 (2,t, J=7.40 Hz), 4.41 (2H, t, J=7.0 Hz), 6.57 (1H, d, J=17.60 Hz), 7.15(5H, m), 7.64 (1, d, J=17.60 Hz), 7.89 (1H, d, J=8.64 Hz), 7.95 (1H, s).MS (m/z): 366 [MH]⁺.

Example 14 Preparation ofN-Hydroxy-3-(2-phenethyl-1-(pyridin-2-yl)methyl-1H-benzimidazol-5-yl)-acrylamide(16)

The titled compound (16) was prepared according to the proceduresdescribed in Example 1, by using appropriate starting materials. HPLC:99.7%; t_(R)=(LC/PDA: Phenomenex Luna C18 2.0×150 mm 5μ column; 0.8mL/min, gradient 5-65% B over 15.5 min, Solvent A: H₂O with 0.1%trifluoroacetic acid; Solvent B: Acetonitrile with 0.1% trifluoroaceticacid; UV 254): 3.11 min. NMR (400 MHz, DMSO-d₆) δ 3.31 (2H, t, J=7.56Hz), 5.81 (2H, s), 6.57 (1H, d, J=17.60 Hz), 7.20-7.36 (6H, m), 7.52(1H, m), 7.64 (1H, d, J=17.60 Hz), 7.68 (1H, d, J=8.48 Hz), 7.77 (1H, d,J=8.48 Hz), 7.87 (1H, m), 8.44 (1H, d, J=3.92 Hz). MS (m/z): 399 [MH]⁺.

Example 15 Preparation ofN-Hydroxy-3-[1-(3-Dimethylamino-2,2-dimethyl-propyl)-2-phenethyl-1H-benzimidazol-5-yl]-acrylamide(17)

The titled compound (17) was prepared according to the proceduresdescribed in Example 1, by using appropriate starting materials. HPLC:100%; t_(R)=(LC/PDA: Phenomenex Luna C18 2.0×150 mm 5μ column; 0.8mL/min, gradient 5-65% B over 15.5 min, Solvent A: H₂O with 0.1%trifluoroacetic acid; Solvent B: Acetonitrile with 0.1% trifluoroaceticacid; UV 254): 2.13 min. NMR (400 MHz, DMSO-d₆) δ 1.08 (6H, s), 2.89(6H, s), 4.30 (2H, s), 6.54 (1H, d, J=15.80 Hz), 7.03 (1H, s), 7.16 (1H,s), 7.22-7.32 (6H, m), 7.65 (1H, d, J=15.80 Hz), 7.91 (1H, s). MS (m/z):421 [MH]⁺.

Example 16 Preparation ofN-Hydroxy-3-[2-Benzyloxymethyl-1-(3-hydroxy-propyl-1H-benzimidazyl-5yl]-acrylamide(19)

The titled compound (19) was prepared according to the proceduresdescribed in Example 1, by using appropriate starting materials. HPLC:98.6%; t_(R)=(LC/PDA: Phenomenex Luna C18 2.0×150 mm 5μ column; 0.8mL/min, gradient 5-65% B over 15.5 min, Solvent A: H₂O with 0.1%trifluoroacetic acid; Solvent B: Acetonitrile with 0.1% trifluoroaceticacid; UV 254): 4.50 min. ¹H NMR (400 MHz, DMSO-d₆) δ 1.94 (2H, m), 3.43(2H, t, J=5.8 Hz), 4.42 (2H, t, J=7.2 Hz), 4.67 (2H, s), 4.97 (2H, s),6.53 (1H, d, J=15.8 Hz), 7.38 (5H, m), 7.63 (1H, d, J=15.8 Hz), 7.67(1H, d, J=9.1 Hz), 7.80 (1H, d, J=8.6 Hz), 7.90 (1H, s), 10.77 (1H, bs).MS (m/z): 382 [MH]⁺.

Example 17 Preparation ofN-Hydroxy-3-[1-(3-hydroxy-propyl)-2-thiophen-3-yl-1H-benzimidazol-5-yl]-acrylamide(20)

The titled compound (20) was prepared according to the proceduresdescribed in Example 1, by using appropriate starting materials. HPLC:97.9%; t_(R)=(LC/PDA: Phenomenex Luna C18 2.0×150 mm 5μ column; 0.8mL/min, gradient 5-65% B over 15.5 min, Solvent A: H₂O with 0.1%trifluoroacetic acid; Solvent B: Acetonitrile with 0.1% trifluoroaceticacid; UV 254): 3.06 min. ¹H NMR (400 MHz, DMSO-d₆), δ 1.98 (2H, m), 3.49(2H, t, J=5.8 Hz), 4.56 (2H, t, J=7.2 Hz), 6.56 (1H, d, J=15.8 Hz), 7.65(1H, d, J=15.8 Hz), 7.69 (1H, d, J=8.7 Hz), 7.75 (1H, dd, J=5.1 Hz, 1.2Hz), 7.89 (2H, m), 7.93 (1H, s), 8.42 (1H, dd, J=2.6 Hz), 10.90 (1H,bs). MS (m/z): 344 [MH]⁺.

Example 18 Preparation ofN-Hydroxy-3-[1-(3-hydroxy-propyl)-2-isobutyl-1H-benzimidazol-5-yl]-acrylamide(21)

The titled compound (21) was prepared according to the proceduresdescribed in Example 1, by using appropriate starting materials. HPLC:100%; t_(R)=(LC/PDA: Phenomenex Luna C18 2.0×150 mm 5μ column; 0.8mL/min, gradient 5-65% B over 15.5 min, Solvent A: H₂O with 0.1%trifluoroacetic acid; Solvent B: Acetonitrile with 0.1% trifluoroaceticacid; UV 254): 3.14 min. ¹H NMR (400 MHz, DMSO-d₆), δ 1.01 (6H, d, J=6.6Hz), 1.94 (2H, m), 2.28 (1H, m), 3.04 (2H, d, J=7.4 Hz), 3.47 (2H, t,J=5.8 Hz), 4.46 (2H, t, J=7.1 Hz), 6.56 (1H, d, J=15.8 Hz), 7.65 (1H, d,J=15.8 Hz), 7.73 (1H, d, J=8.6 Hz), 7.89 (1H, d, J=8.6 Hz), 7.94 (1H,s). MS (m/z): 318 [MH]⁺.

Example 19 Preparation ofN-Hydroxy-3-[1-(3-hydroxy-propyl)-2-octyl-1H-benzimidazol-5-yl]-acrylamide(23)

The titled compound (23) was prepared according to the proceduresdescribed in Example 1, by using appropriate starting materials. HPLC:99.0%; t_(R)=(LC/PDA: Phenomenex Luna C18 2.0×150 mm 5μ column; 0.8mL/min, gradient 5-65% B over 15.5 min, Solvent A: H₂O with 0.1%trifluoroacetic acid; Solvent B: Acetonitrile with 0.1% trifluoroaceticacid; UV 254): 7.38 min. ¹H NMR (400 MHz, DMSO-d₆) δ 0.86 (3H, t, J=6.8Hz), 1.32 (10H, m), 1.83 (2H, m), 1.94 (2H, m), 3.12 (2H, t, J=7.7 Hz),3.46 (2H, t, J=5.8 Hz), 4.44 (2H, t, J=7.0 Hz), 6.56 (1H, d, J=15.8 Hz),7.64 (1H, d, J=15.8 Hz), 7.71 (1H, d, J=8.6 Hz), 7.87 (1H, d, J=8.6 Hz),7.92 (1H, s). MS (m/z): 374 [MH]⁺.

Example 20 Preparation ofN-Hydroxy-[2-cyclohexyl-1-(3-hydroxy-propyl)-1H-benzimidazol-5-yl]-acrylamide(24)

The titled compound (24) was prepared according to the proceduresdescribed in Example 1, by using appropriate starting materials. HPLC:98.0%; t_(R)=(LC/PDA: Phenomenex Luna C18 2.0×150 mm 5μ column; 0.8mL/min, gradient 5-65% B over 15.5 min, Solvent A: H₂O with 0.1%trifluoroacetic acid; Solvent B: Acetonitrile with 0.1% trifluoroaceticacid; UV 254): 7.38 min. ¹H NMR (400 MHz, DMSO-d₆) δ 1.28-2.03 (12H, m),3.33 (1H, m), 3.47 (2H, t, J=5.7 Hz), 4.51 (2H, t, J=6.9 Hz), 6.58 (1H,d, J=15.8 Hz), 7.65 (1H, d, J=15.8 Hz), 7.76 (1H, d, J=8.6 Hz), 7.92(1H, d, J=8.7 Hz), 7.93 (1H, s), 10.85 (1H, bs). MS (m/z): 344 [MH]⁺.

Example 21 Preparation ofN-Hydroxy-3-(2-isobutyl-1-phenethyl-1H-benzimidazol-5-yl]-acrylamide(25)

The titled compound (25) was prepared according to the proceduresdescribed in Example 1, by using appropriate starting materials. HPLC:99.1%; t_(R)=(LC/PDA: Phenomenex Luna C18 2.0×150 mm 5μ column; 0.8mL/min, gradient 5-65% B over 15.5 min, Solvent A: H₂O with 0.1%trifluoroacetic acid; Solvent B: Acetonitrile with 0.1% trifluoroaceticacid; UV 254): 6.51 min. ¹H NMR (400 MHz, DMSO-d₆) 80.90 (6H, d, J=6.6Hz), 2.10 (1H, m), 2.70 (2H, d, J=7.3 Hz), 3.11 (2H, t, J=7.0 Hz), 4.66(2H, t, J=7.0 Hz), 6.57 (1H, d, J=15.8 Hz), 7.14 (2H, m), 7.26 (3H, m),7.64 (1H, d, J=15.8 Hz), 7.70 (1H, d, J=8.8 Hz), 7.86 (1H, d, J=8.6 Hz),7.92 (1H, s); ¹³C NMR (100 MHz, DMSO-d₆) δ 22.0, 26.9, 33.3, 34.5, 45.8,113.0, 114.3, 119.7, 123.7, 126.9, 128.5, 129.0, 132.2, 132.7, 137.2,137.8, 154.4, 162.5. MS (m/z): 364 [MH]⁺.

Example 22 Preparation ofN-Hydroxy-3-(1,2-Diphenethyl-1H-benzimidazol-5-yl]-acrylamide (26)

The titled compound (26) was prepared according to the proceduresdescribed in Example 1, by using appropriate starting materials. HPLC:98.3%; t_(R)=(LC/PDA: Phenomenex Luna C18 2.0×150 mm 5μ column; 0.8mL/min, gradient 5-65% B over 15.5 min, Solvent A: H₂O with 0.1%trifluoroacetic acid; Solvent B: Acetonitrile with 0.1% trifluoroaceticacid; UV 254): 7.68 min. ¹H NMR (400 MHz, DMSO-d₆) δ 2.99 (4H, m), 3.09(2H, m), 4.59 (2H, t, J=6.9 Hz), 6.56 (1H, d, J=15.8 Hz), 7.07 (2H, m),7.23 (6H, m), 7.31 (2H, m), 7.64 (1H, d, J=15.5 Hz), 7.66 (1H, d, J=7.2Hz), 7.78 (1H, d, J=8.6 Hz), 7.92 (1H, s); ¹³C NMR (100 MHz, DMSO-d₆) δ27.0, 31.9, 34.5, 45.6, 112.7, 114.7, 119.4, 123.5, 126.5, 126.9, 128.3,128.5, 129.0, 131.8, 133.0, 137.3, 138.0, 139.5, 154.6, 162.6. MS (m/z):412 [MH]⁺.

Example 23 Preparation ofN-Hydroxy-3-(2-phenethyl-1-(2-pyridin-3-yl-ethyl)-1H-benzimidazol-5-yl]-acrylamide(27)

The titled compound (27) was prepared according to the proceduresdescribed in Example 1, by using appropriate starting materials. HPLC:99.9%; t_(R)=(LC/PDA: Phenomenex Luna C18 2.0×150 mm 5μ column; 0.8mL/min, gradient 5-65% B over 15.5 min, Solvent A: H₂O with 0.1%trifluoroacetic acid; Solvent B: Acetonitrile with 0.1% trifluoroaceticacid; UV 254): 3.42 min. ¹H NMR (400 MHz, DMSO-d₆) δ 3.10 (4H, m), 3.28(2H, t), 4.63 (2H, t) 6.53 (1H, d), 7.22-7.33 (7H, m), 7.54-7.74 (4H,m), 8.55 (2H, d), 10.88 (1H, bs). MS (m/z): 413 [MH]⁺.

Example 24 Preparation ofN-Hydroxy-3-[1-(3-Hydroxy-propyl)-2-isobutyl-1H-benzimidazol-5-yl]-propionamide(29)

The titled compound (29) was prepared according to the proceduresdescribed in Example 1, by using appropriate starting materials. HPLC:99.6%; t_(R)=(LC/PDA: Phenomenex Luna C18 2.0×150 mm 5μ column; 0.8mL/min, gradient 5-65% B over 15.5 min, Solvent A: H₂O with 0.1%trifluoroacetic acid; Solvent B: Acetonitrile with 0.1% trifluoroaceticacid; UV 254): 2.88 min. ¹H NMR (400 MHz, DMSO-d₆) δ 1.00 (6H, d, J=6.4Hz), 2.06 (2H, m), 2.27 (1H, m), 2.42 (2H, t, J=7.6 Hz), 3.05-3.11 (4H,m), 3.57 (2H, t, J=6.0 Hz), 4.52 (2H, t, J=7.2 Hz), 7.45 (1H, d, J=8.0Hz), 7.56 (1H, s), 7.78 (1H, d, J=8.0 Hz); ¹³C NMR (100 MHz, MeOD) δ20.6 (2C), 27.2, 30.4, 30.6, 32.7, 33.5, 41.5, 57.0, 112.0, 112.3,112.4, 126.3, 129.9, 139.6, 152.3, 169.4. MS (m/z): 320 [MH]J.

Example 25 Preparation ofN-Hydroxy-3-{1-[3-(2-oxo-pyrrolidin-1-yl)-propyl]-2-phenethyl-1H-benzimidazol-5-yl}-acrylamide(30)

The titled compound (30) was prepared according to the proceduresdescribed in Example 1, by using appropriate starting materials. HPLC:99.7%; t_(R)=(LC/PDA: Phenomenex Luna C18 2.0×150 mm 5μ column; 0.8mL/min, gradient 5-65% B over 15.5 min, Solvent A: H₂O with 0.1%trifluoroacetic acid; Solvent B: Acetonitrile with 0.1% trifluoroaceticacid; UV 254): 2.88 min. ¹H NMR (400 MHz, DMSO-d₆) δ 1.84 (4H, m),3.14-3.41 (8H, m), 4.29 (2H, t, J=7.04 Hz), 6.54 (1H, d, J=15.76 Hz),7.21-7.33 (5H, m), 7.62 (1H, d, J=15.76 Hz), 7.71 (1H, d, J=8.36 Hz),7.84 (1H, d, J=8.36 Hz), 7.93 (1H, s). MS (m/z): 433 [MH]⁺.

Example 26 Preparation ofN-Hydroxy-3-[1-(3-morpholin-4-propyl]-2-phenethyl-1H-benzimidazol-5-yl}-acrylamide(31)

The titled compound (31) was prepared according to the proceduresdescribed in Example 1, by using appropriate starting materials. HPLC:99.7%; t_(R)=(LC/PDA: Phenomenex Luna C18 2.0×150 mm 5μ column; 0.8mL/min, gradient 5-65% B over 15.5 min, Solvent A: H₂O with 0.1%trifluoroacetic acid; Solvent B: Acetonitrile with 0.1% trifluoroaceticacid; UV 254): 2.16 min. ¹H NMR (400 MHz, DMSO-d₆) δ 2.12 (2H, m), 3.11(6H, m), 3.39 (2H, t, J=7.44 Hz), 4.39 (2H, t, J=7.01 Hz), 6.56 (1H, d,J=15.8 Hz), 7.23-7.33 (5H, m), 7.62 (1H, d, J=15.8 Hz), 7.71 (1H, d,J=8.60 Hz), 7.85 (1H, d, J=8.60 Hz), 7.95 (1H, s). MS (m/z): 435 [MH]⁺.

Example 27 Preparation of3-[5-(2-Hydrocarbamoyl-vinyl)-2-phenethyl-benzimidazol-1-yl]-propionicacid (32)

The titled compound (32) was prepared according to the proceduresdescribed in Example 1, by using appropriate starting materials. HPLC:95.6%; t_(R)=(LC/PDA: Phenomenex Luna C18 2.0×150 mm 5μ column; 0.8mL/min, gradient 5-65% B over 15.5 min, Solvent A: H₂O with 0.1%trifluoroacetic acid; Solvent B: Acetonitrile with 0.1% trifluoroaceticacid; UV 254): 2.55 min. ¹H NMR (400 MHz, DMSO-d₆) δ 2.74 (2H, t, J=6.68Hz), 4.49 (2H, t, J=6.68 Hz), 3.16 (2H, t, J=7.44 Hz), 6.52 (1H, d,J=15.76 Hz), 7.22-7.33 (5H, m), 7.62 (1H, d, J=15.76 Hz), 7.66 (1H, d,J=8.56 Hz), 7.82 (1H, d, J=8.56 Hz), 7.89 (1H, s), 11.00 (1H, s) MS(m/z): 380 [MH]⁺.

Example 28 Preparation ofN-Hydroxy-3-(1-Benzyl-2-phenethyl-1H-benzimidazol-5-yl}-acrylamide (33)

The titled compound (33) was prepared according to the proceduresdescribed in Example 1, by using appropriate starting materials. HPLC:99.0%; t_(R)=(LC/PDA: Phenomenex Luna C18 2.0×150 mm 5μ column; 0.8mL/min, gradient 5-65% B over 15.5 min, Solvent A: H₂O with 0.1%trifluoroacetic acid; Solvent B: Acetonitrile with 0.1% trifluoroaceticacid; UV 254): 7.82 min. ¹H NMR (400 MHz, DMSO-d₆) δ 3.08 (2H, t, J=7.4Hz), 3.34 (2H, t, J=7.5 Hz), 5.62 (2H, s), 6.50 (1H, d, J=15.8 Hz), 7.14(2H, m), 7.30 (8H, m), 7.63 (3H, m), 7.92 (1H, s), 10.78 (1H, br); ¹³CNMR (100 MHz, DMSO-d₆) δ 27.8, 32.2, 46.8, 112.1, 115.9, 118.6, 123.0,126.4, 126.8, 127.9, 128.3, 128.4, 128.9, 131.0, 134.4, 135.7, 138.4,139.9, 155.3, 162.7. MS (m/z): 398 [MH]⁺.

Example 29 Preparation ofN-Hydroxy-3-(1-Benzyl-2-isobutyl-1H-benzimidazol-5-yl}-acrylamide (34)

The titled compound (34) was prepared according to the proceduresdescribed in Example 1, by using appropriate starting materials. HPLC:89.2%; t_(R)=(LC/PDA: Phenomenex Luna C18 2.0×150 mm 5μ column; 0.8mL/min, gradient 5-65% B over 15.5 min, Solvent A: H₂O with 0.1%trifluoroacetic acid; Solvent B: Acetonitrile with 0.1% trifluoroaceticacid; UV 254): 6.07 min. ¹H NMR (400 MHz, CDCl₃) δ 0.92 (6H, d, J=6.6Hz), 2.13 (1H, m), 3.02 (2H, d, J=7.4 Hz), 5.72 (2H, s), 6.54 (1H, d,J=15.8 Hz), 7.21 (2H, m), 7.35 (3H, m), 7.66 (3H, m), 7.96 (1H, s); ¹³CNMR (100 MHz, CDCl₃) δ 22.0, 27.2, 34.0, 47.2, 112.8, 114.9, 119.4,123.7, 126.8, 128.0, 128.9, 131.9, 133.6, 135.3, 138.0, 155.0, 162.6. MS(m/z): 350 [MH]⁺.

Example 30 Preparation ofN-Hydroxy-3-(1-Benzyl-1H-benzimidazol-5-yl}-acrylamide (35)

The titled compound (35) was prepared according to the proceduresdescribed in Example 1, by using appropriate starting materials. HPLC:97.0%; t_(R)=(LC/PDA: Phenomenex Luna C18 2.0×150 mm 5μ column; 0.8mL/min, gradient 5-65% B over 15.5 min, Solvent A: H₂O with 0.1%trifluoroacetic acid; Solvent B: Acetonitrile with 0.1% trifluoroaceticacid; UV 254): 3.69 min. ¹H NMR (400 MHz, CD₃OD) δ 5.68 (2H, s), 6.54(1H, d, J=15.7 Hz), 7.37 (5H, m), 7.66 (1H, d, J=15.8 Hz), 7.75 (2H, s),7.94 (1H, s), 9.36 (1H, br); ¹³C NMR (100 MHz, CD₃OD) δ 51.7, 114.8,116.1, 120.6, 126.5, 129.2, 130.2, 130.4, 135.0, 135.3, 140.1, 165.6. MS(m/z): 294 [MH]⁺.

Example 31 Preparation ofN-Hydroxy-3-(2-phenethyl-1-propyl-1H-benzimidazol-5-yl}-acrylamide (36)

The titled compound (36) was prepared according to the proceduresdescribed in Example 1, by using appropriate starting materials. HPLC:93.9%; t_(R)=(LC/PDA: Phenomenex Luna C18 2.0×150 mm 5μ column; 0.8mL/min, gradient 5-65% B over 15.5 min, Solvent A: H₂O with 0.1%trifluoroacetic acid; Solvent B: Acetonitrile with 0.1% trifluoroaceticacid; UV 254): 6.05 min. ¹H NMR (400 MHz, CD₃OD) δ 0.90 (3H, t, J=7.4Hz), 1.70 (2H, m), 3.20 (2H, m), 3.48 (2H, t, J=7.1 Hz), 4.21 (2H, t,J=7.4 Hz), 6.54 (1H, d, J=15.7 Hz), 7.20 (5H, m), 7.65 (1H, d, J=15.7Hz), 7.75 (1H, d, J=8.8 Hz), 7.79 (1H, d, J=8.6 Hz), 7.84 (1H, s); ¹³CNMR (100 MHz, CD₃OD) δ 11.2, 23.6, 28.7, 34.0, 47.7, 114.4, 114.6,120.5, 126.3, 128.3, 129.5, 130.0, 132.7, 134.0, 135.2, 139.9, 140.1,155.5, 165.6. MS (m/z): 350 [MH]⁺.

Example 32 Preparation ofN-Hydroxy-3-(1-propyl-1H-benzimidazol-5-yl}-acrylamide (37)

The titled compound (37) was prepared according to the proceduresdescribed in Example 1, by using appropriate starting materials. HPLC:95.2%; t_(R)=(LC/PDA: Phenomenex Luna C18 2.0×150 mm 5μ column; 0.8ml/min, gradient 5-65% B over 15.5 min, Solvent A: H₂O with 0.1%trifluoroacetic acid; Solvent B: Acetonitrile with 0.1% trifluoroaceticacid; UV 254): 2.92 min. ¹H NMR (400 MHz, CD₃OD) δ 0.97 (3H, t, J=7.4Hz), 1.98 (2H, m), 4.42 (2H, t, J=7.3 Hz), 6.55 (1H, d, J=15.8 Hz), 7.68(1H, d, J=15.8 Hz), 7.79 (1H, d, J=8.7 Hz), 7.88 (1H, d, J=8.7 Hz), 7.92(1H, s), 9.24 (1H, s); ¹³C NMR (100 MHz, CD₃CD) δ 11.1, 23.8, 48.4,114.3, 116.1, 120.3, 126.4, 133.8, 134.9, 135.0, 140.3, 143.5, 165.7. MS(m/z): 246 [MH]⁺.

Example 33 Preparation ofN-Hydroxy-3-(1-Ethyl-2-phenethyl-1H-benzimidazol-5-yl}-acrylamide (38)

The titled compound (38) was prepared according to the proceduresdescribed in Example 1, by using appropriate starting materials. HPLC:99.0%; t_(R)=(LC/PDA: Phenomenex Luna C18 2.0×150 mm 5μ column; 0.8mL/min, gradient 5-65% B over 15.5 min, Solvent A: H₂O with 0.1%trifluoroacetic acid; Solvent B: Acetonitrile with 0.1% trifluoroaceticacid; UV 254): 5.06 min. ¹H NMR: (400 MHz, CD₃OD) δ 1.37 (3H, t, J=7.3Hz), 3.26 (2H, t, J=7.6 Hz), 3.53 (2H, t, J=7.5 Hz), 4.78 (2H, dd, J=7.3Hz), 6.60 (1H, d, J=15.8 Hz), 7.21-7.31 (5H, m), 7.72 (1H, d, J=15.8Hz), 7.83-7.89 (3H, m). MS (m/z): 336 [MH]⁺.

Example 34 Preparation ofN-Hydroxy-3-(1-Ethyl-H-benzimidazol-5-yl)-acrylamide (39)

The titled compound (39) was prepared according to the proceduresdescribed in Example 1, by using appropriate starting materials. HPLC:99.0%; t_(R)=(LC/PDA: Phenomenex Luna C18 2.0×150 mm 5μ column; 0.8ml/min, gradient 5-65% B over 15.5 min, Solvent A: H₂O with 0.1%trifluoroacetic acid; Solvent B: Acetonitrile with 0.1% trifluoroaceticacid; UV 254): 1.86 min. ¹H NMR: (400 MHz, CD₃OD) δ 1.64 (3H, t, J=7.3Hz), 4.55 (2H, dd, J=7.3 Hz), 6.61 (1H, d, J=15.8 Hz), 7.72 (1H, d,J=15.8 Hz), 7.86-7.97 (3H, m), 9.38 (1H, s). MS (m/z): 232 [MH]⁺.

Example 35 Preparation of1-(3-Hydroxy-propyl)-2-phenethyl-1H-benzimidazol-5-carboxylic acidhydroxyamide (40)

The title compound (40) was prepared according to the proceduresdescribed in Example 1, by using appropriate starting materials. HPLC:96.0%. ¹H NMR (400 MHz, CD3OD, δ): 1.88 (2H, m), 3.16 (2H, t, J=7.2 Hz),3.46 (4H, m), 4.34 (2H, t, J=7.2 Hz), 7.12-7.21 (5H, m), 7.82 (2H, m),8.05 (1H, s). MS (m/z): 340 [MH]⁺.

Example 36 Preparation ofN-Hydroxy-3-[1-(2-pyridin-2-yl-ethyl)-1H-benzimidazol-5-yl]-acrylamide(42)

The titled compound (42) was prepared according to the proceduresdescribed in Example 1, by using appropriate starting materials. HPLC:98.4%; t_(R)=(LC/PDA: Phenomenex Luna C18 2.0×150 mm 5μ column; 0.8mL/min, gradient 5-65% B over 15.5 min, Solvent A: H₂O O with 0.1%trifluoroacetic acid; Solvent B: Acetonitrile with 0.1% trifluoroaceticacid; UV 254): 1.05 min. ¹H NMR (400 MHz, DMSO-d₆) δ 3.43 (2H, t), 4.84(2H, t), 6.53 (1H, d), 7.41 (2H, m), 7.64 (2H, m), 7.77-7.95 (4H, m),8.56 (1H, s), 9.16 (1H, s). MS (m/z): 309 [MH]⁺.

Example 37 Preparation ofN-Hydroxy-3-(1-Ethyl-2-methyl-1H-benzimidazol-5-yl]-acrylamide (43)

The titled compound (43) was prepared according to the proceduresdescribed in Example 1, by using appropriate starting materials. HPLC:96.5%; t_(R)=(LC/PDA: Phenomenex Luna C18 2.0×150 mm 5μ column; 0.8mL/min, gradient 5-65% B over 15.5 min, Solvent A: H₂O with 0.1%trifluoroacetic acid; Solvent B: Acetonitrile with 0.1% trifluoroaceticacid; UV 254): 2.52 min. ¹H NMR (400 MHz, DMSO-d₆) δ 1.38 (3H, t), 2.85(3H, s), 4.42 (2H, t), 6.58 (1H, d), 7.31 (1H, m), 7.50 (1H, d), 7.88(2H, m), 10.31 (1H, bs), 11.18 (1H, bs). MS (m/z): 246 [MH]⁺.

Example 38 Preparation ofN-Hydroxy-3[1-(3-hydroxy-propyl)-1H-benzimidazol-5-yl]-acrylamide (47)

The titled compound (47) was prepared according to the proceduresdescribed in Example 1, by using appropriate starting materials.HPLC: >99%; t_(R)=(LC/PDA: Phenomenex Luna C18 2.0×150 mm 5μ column; 0.8mL/min, gradient 5-65% B over 15.5 min, Solvent A: H₂O with 0.1%trifluoroacetic acid; Solvent B: Acetonitrile with 0.1% trifluoroaceticacid; UV 254): 1.02 min. ¹H NMR (400 MHz, CD₃OD) δ 2.12 (2H, m), 3.58(2H, t, J=5.7 Hz), 4.57 (2H, t, J=6.9 Hz), 6.55 (1H, d, J=15.8 Hz), 7.67(1H, d, J=15.8 Hz), 7.79 (1H, d, J=8.7 Hz), 7.89 (1H, d, J=8.9 Hz), 7.92(1H, s), 9.22 (1H, s); ¹³C NMR (100 MHz, MeOD) δ 32.7, 45.3, 59.2,114.3, 116.1, 120.3, 126.4, 135.0, 140.3, 143.8, 165.7. MS (m/z): 262[MH]⁺.

Example 39 Preparation ofN-Hydroxy-3-(1-methyl-2-phenethyl-1H-benzimidazol-5-yl)-acrylamide (48)

The titled compound (48) was prepared according to the proceduresdescribed in Example 1, by using appropriate starting materials. HPLC:99%; t_(R)=(LC/PDA: Phenomenex Luna C18 2.0×150 mm 5μ column; 0.8mL/min, gradient 5-65% B over 15.5 min, Solvent A: H₂O with 0.1%trifluoroacetic acid; Solvent B: Acetonitrile with 0.1% trifluoroaceticacid; UV 254): 4.53 min. ¹H NMR: (400 MHz, CD₃OD) δ 3.18 (2H, t, J=7.5Hz), 3.47 (2H, t, J=7.4 Hz), 3.76 (3H, s), 6.54 (1H, d, J=15.8 Hz),7.10-7.26 (5H, m), 7.65 (1H, d, J=15.8 Hz), 7.75-7.82 (3H, m). MS (m/z):322 [MH]⁺.

Example 40 Preparation ofN-Hydroxy-3-(2-phenethyl-1H-benzimidazol-5-yl)-acrylamide (50)

The titled compound (50) was prepared according to the proceduresdescribed in Example 1, by using appropriate starting materials. HPLC:99%; t_(R)=(LC/PDA: Phenomenex Luna C18 2.0×150 mm 5μ column; 0.8mL/min, gradient 5-65% B over 15.5 min, Solvent A: H₂O with 0.1%trifluoroacetic acid; Solvent B: Acetonitrile with 0.1% trifluoroaceticacid; UV 254): 4.36 min. ¹H NMR (400 MHz, DMSO-d₆) δ 3.16 (2H, t, J=7.5Hz), 3.36 (2H, t, J=7.9 Hz), 6.53 (1H, d, J=15.8 Hz), 7.17-7.29 (5H, m),7.58 (1H, d, J=15.8 Hz), 7.66-7.87 (3H, m). MS (m/z): 308 [MH]⁺.

Example 41 Preparation of N-Hydroxy-3-(1H-benzimidazol-5-yl)-acrylamide(51)

The titled compound (51) was prepared according to the proceduresdescribed in Example 1, by using appropriate starting materials. HPLC:99%; t_(R)=(LC/PDA: Phenomenex Luna C18 2.0×150 mm 5μ column; 0.8mL/min, gradient 5-65% B over 15.5 min, Solvent A: H₂O with 0.1%trifluoroacetic acid; Solvent B: Acetonitrile with 0.1% trifluoroaceticacid; UV 254): 0.99 min. ¹H NMR (400 MHz, DMSO-d₆) δ 6.62 (1H, d, J=15.8Hz), 7.74 (1H, d, J=15.8 Hz), 7.85-7.99 (3H, m), 9.32 (1H, s). MS (m/z):204 [MH]⁺.

Example 42 Preparation ofN-Hydroxy-3-[1-methyl-2-(3-phenyl-propyl)-1H-benzimidazol-5-yl]-acrylamide(52) Step 1

To a pre-stirred solution of methyltrans-4-(methylamine)-3-nitrocinnamate (1.0 g, 4.0 mmol; prepared asdescribed in Example 1) in 40 mL methanol and 10 mL glacial acetic acid,was added Tin chloride (3.0 g, 16.0 mmol). The resulting solution washeated to 55° C. for 24 hours and then cooled to room temperature. Thesolvent was removed and the mixture was neutralized with sodiumbicarbonate to pH=8. The crude product was extracted withdichloromethane (20 mL) for three times. The organic extracts werecombined and washed with water (10 mL) twice and brine (10 mL) once andfurther dried over Na₂SO₄ for 1 hour, filtered and concentrated. Theproduct methyl trans-4-(methylamine)-3-aminocinnamate was obtained in82.5% yield (726 mg). MS(m/z): 207 [MH]⁺.

Step 2

4-phenylbutyric acid (68 mg, 0.41 mmol), methyltrans-4-(methylamine)-3-aminocinnamate (85 mg, 0.40 mmol) and PyBOP (236mg, 0.46 mmol) were mixed in a 25 mL round bottom flask with 10 mL ofdried dichloromethane. The resulting mixture was stirred under nitrogenatmosphere for 5 minutes. DIEA (288 uL, 1.62 mmol) was injected and theresulting mixture was stirred at room temperature for another 4 hours.The progress of the reaction was monitored by TLC. The couplingproducts, 3-{3-amino-4-[methyl-(4-phenyl-butyryl)amino]-phenyl}-acrylicacid methyl ester and3-[4-methylamino-3-(4-phenyl-butyrylamino)-phenyl]-acrylic acid methylester, were obtained (110 mg 78%) after purification using columnchromatography. (Solvent system: Ethyl acetate:hexane=1:1) MS(m/z): 353[MH]J.

Step 3

The above coupling products (59 mg, 0.17 mmol) was heated with 5 mL ofglacial acetic acid at 70° C. for 4 hours. After cooling down to roomtemperature, the pure product,3-[1-methyl-2-(3-phenyl-propyl)-1H-benzimidazol-5-yl]-acrylic acidmethyl ester, was obtained quantitatively by removing glacial aceticacid under vacuum. ¹H NMR (400 MHz, DMSO-d₆) δ 2.14 (2H, m), 2.75 (2H,t), 3.14 (2H, t), 3.95 (3H, s), 6.58 (1H, d), 7.16-7.30 (5H, m), 7.65(1H, d), 7.72 (1H, d), 7.90 (2H, m). MS(m/z): 335 [MH]⁺.

Step 4

The titled compound (52) was prepared according to the procedures forpreparation of hydroxamic acid as described in Example 1, by usingappropriate starting materials. HPLC: 99.8%; t_(R)=(LC/PDA: PhenomenexLuna C18 2.0×150 mm 5μ column; 0.8 mL/min, gradient 5-65% B over 15.5min, Solvent A: H₂O with 0.1% trifluoroacetic acid; Solvent B:Acetonitrile with 0.1% trifluoroacetic acid; UV 254): 5.01 min. ¹H NMR(400 MHz, DMSO-d₆) δ 2.14 (2H, m), 2.75 (2H, t), 3.14 (2H, t), 3.95 (3H,s), 6.58 (1H, d), 7.16-7.30 (5H, m), 7.65 (1H, d), 7.72 (1H, d), 7.90(2H, m), 10.89 (1H, bs). MS (m/z): 336 [MH]⁺.

Example 43 Preparation ofN-Hydroxy-3-[1-(3-imidazol-1-yl-propyl)-2-phenethyl-1H-benzimidazol-5-yl]-acrylamide(56)

The titled compound (56) was prepared according to the proceduresdescribed in Example 1, by using appropriate starting materials. HPLC:98.0%; t_(R)=(LC/PDA: Phenomenex Luna C18 2.0×150 mm 5μ column; 0.8mL/min, gradient 5-65% B over 15.5 min, Solvent A: H₂O with 0.1%trifluoroacetic acid; Solvent B: Acetonitrile with 0.1% trifluoroaceticacid; UV 254): 3.50 min. ¹H NMR (400 MHz, CD₃OD) δ 2.20 (2H, m), 3.19(2H, m), 3.39 (2H, t, J=7.6 Hz), 4.28 (4H, t, J=7.6 Hz), 6.52 (1H, d,J=16.0 Hz), 7.17 (5H, m), 7.52 (1H, t, J=1.5 Hz), 7.58 (1H, t, J=1.6Hz), 7.65 (1H, d, J=16.0 Hz), 7.68 (2H, s), 7.85 (1H, s), 8.84 (1H, s);¹³C NMR (100 MHz, CD₃OD) δ 29.3, 30.7, 34.4, 42.4, 47.6, 113.0, 116.2,119.2, 121.6, 123.1, 125.7, 128.0, 129.6, 129.9, 133.7, 135.1, 136.6,137.2, 140.7, 140.9, 156.5, 166.0. MS (m/z): 416 [MH]⁺.

Example 44 Preparation ofN-Hydroxy-3-[1-(4-dimethylamino-butyl)-2-phenethyl-1H-benzimidazol-5-yl]-acrylamide(57)

The titled compound (57) was prepared according to the proceduresdescribed in Example 1, by using appropriate starting materials. HPLC:97.0%; t_(R)=(LC/PDA: Phenomenex Luna C18 2.0×150 mm 5μ column; 0.8mL/min, gradient 5-65% B over 15.5 min, Solvent A: H₂O with 0.1%trifluoroacetic acid; Solvent B: Acetonitrile with 0.1% trifluoroaceticacid; UV 254): 3.70 min. ¹H NMR (400 MHz, CD₃OD) δ 1.71 (4H, m), 2.82(6H, s), 3.05 (2H, t, J=7.1 Hz), 3.21 (2H, t, J=7.6 Hz), 3.44 (2H, t,J=7.5 Hz), 4.27 (2H, t, J=7.5 Hz), 6.53 (1H, d, J=16.0 Hz), 7.20 (5H,m), 7.65 (1H, d, J=16.0 Hz), 7.73 (2H, m), 7.85 (1H, s); ¹³C NMR (100MHz, CD₃OD) δ 22.8, 27.3, 29.1, 34.2, 43.5, 45.1, 58.3, 113.5, 115.6,119.6, 125.9, 128.1, 129.5, 130.0, 134.2, 134.7, 140.4, 140.6, 156.2,162.7, 165.9. MS (m/z): 407 [MH]⁺.

Example 45 Preparation ofN-Hydroxy-3-[1-(3-Hydroxy-propyl)-2-isobutyl-1H-benzimidazol-5-yl]-acrylamide(29) Step 1

3-[1-(3-hydroxy-propyl)-2-isobutyl-1H-benzimidazol-5-yl]-acrylic acidmethyl ester (prepared according to the Example 1, step 1-3) (126.6 mg,0.4 mmol) and 10% Pd/C (40 mg) in 10 mL of MeOH was hydrogenated using ahydrogen balloon overnight. After filtration through short-column silicagel, the filtrate was evaporated under reduced pressure to give3-[1-(3-hydroxy-propyl)-2-isobutyl-1H-benzimidazol-5-yl]-propionic acidmethyl ester (127 mg) in quantitative yield: MS m/z (M+H)+: 319; ¹H NMR(400 MHz, MeOD) δ 0.95 (6H, d, J=6.4 Hz), 1.92 (2H, m), 2.19 (1H, m),2.60 (2H, t, J=8.0 Hz), 2.74 (2H, d, J=7.2 Hz), 2.96 (2H, t, J=7.6 Hz),3.50 (2H, t, J=4.1 Hz), 3.54 (3H, s), 4.25 (2H, t, J=7.2 Hz), 7.05 (1H,d, J=8.0 Hz), 7.30-7.40 (2H, m); ¹³C NMR (100 MHz, MeOD) δ 20.9 (2C),27.3, 30.1, 31.5, 34.6, 35.3, 39.5, 50.1, 57.4, 109.1, 116.4, 122.1,132.6, 134.2, 141.3, 154.2, 173.2.

Step 2

The titled compound (29) was prepared according the method describedpreviously for the preparation of hydroxamic acid: MS m/z (M+H)⁺: 320;¹H NMR (400 MHz, MeOD) δ 1.00 (6H, d, J=6.4 Hz), 2.06 (2H, m), 2.27 (1H,m), 2.42 (2H, t, J=7.6 Hz), 3.05-3.11 (4H, m), 3.57 (2H, t, J=6.0 Hz),4.52 (2H, t, J=7.2 Hz), 7.45 (1H, d, J=8.0 Hz), 7.56 (1H, s), 7.78 (1H,d, J=8.0 Hz); ¹³C NMR (100 MHz, MeOD) δ 20.6 (2C), 27.2, 30.4, 30.6,32.7, 33.5, 41.5, 57.0, 112.0, 112.3, 112.4, 126.3, 129.9, 139.6, 152.3,169.4.

Example 46 Preparation ofN-Hydroxy-3-[2-(benzylamino-methyl)-1-methyl-1H-benzimidazol-5-yl]-acrylamide(60) Step 1

3-[2-(N-Fmoc-aminomethyl)-1methyl-1H-benzimidazol-5-yl]-acrylic acidmethyl ester (43 mg, 0.176 mmol, prepared according to Example 42, step1-3 by using appropriate starting materials) was dissolved in 10 mL ofdichloromethane. The resulting solution was treated with 2.0 mL ofpiperidine. Removed all the solvent and piperidine under vacuum gave3-(2-aminomethyl-1-methyl-1H-benzimidazol-5-yl)-acrylic acid methylester. MS(m/z): 246 [MH]⁺.

Step 2

Benzaldehyde (47 mg, 0.445 mmol),3-(2-aminomethyl-1-methyl-1H-benzimidazol-5-yl)-acrylic acid methylester (109 mg, 80%, 0.445 mmol) and acetic acid (27 mg, 0.445 mmol) weredissolved in 15 mL of dichloromethane. The mixture was stirred at roomtemperature for 1 hour. Sodium triacetoxyborohydride (142 mg, 95%, 0.668mmol) was added to the above solution. The reaction was completed after12 hours and the organic layer was washed with saturated NaHCO₃ (10 mL)twice, followed by washing with water (10 mL) twice, with brine (10 mL)once and then dried over Na₂SO₄. After filtration, the crude product(100 mg, 67.6% yield),3-[2-(benzylamino-methyl)-1-methyl-1H-bezimidazol-5-yl]-acrylic acidmethyl ester, was obtained by removing the solvent. MS(m/z): 336 [MH]⁺.

Step 3

The titled compound (60) was prepared according to the proceduresdescribed in Step 4 of Example 1, by using3-[2-(benzylamino-methyl)-1-methyl-1H-bezimidazol-5-yl]-acrylic acidmethyl ester as the starting material HPLC: 89.6%; t_(R)=(LC/PDA:Phenomenex Luna C18 2.0×150 mm 5μ column; 0.8 mL/min, gradient 5-65% Bover 15.5 min, Solvent A: H₂O with 0.1% trifluoroacetic acid; Solvent B:Acetonitrile with 0.1% trifluoroacetic acid; UV 254): 3.68 min. ¹H NMR(400 MHz, DMSO-d₆) δ 3.78 (3H, s), 4.37 (2H, s), 4.58 (2H, s), 6.48 (1H,d), 7.46 (3H, m), 7.55 (3H, m), 7.64 (2H, t), 7.88 (1H, s), 9.88 (1H,bs), 10.74 (1H, bs). MS (m/z): 337 [MH]⁺.

Example 47 Preparation ofN-Hydroxy-3-[1-(3-dimethylamino-propyl)-2-phenethyl-1H-benzimidazol-5-yl]-acrylamide(63)

The titled compound (63) was prepared according to the proceduresdescribed in Example 1, by using appropriate starting materials. HPLC:100%; t_(R)=(LC/PDA: Phenomenex Luna C18 2.0×150 mm 5μ column; 0.8mL/min, gradient 5-65% B over 15.5 min, Solvent A: H₂O with 0.1%trifluoroacetic acid; Solvent B: Acetonitrile with 0.1% trifluoroaceticacid; UV 254): 3.52 min. ¹H NMR (400 MHz, DMSO-d₆) δ 2.09 (2H, m), 2.75(3H, s), 2.76 (3H, s), 3.12-3.22 (4H, m), 3.37 (2H, b), 4.50 (2H, b),6.55 (1H, d, J=15.76 Hz), 7.22-7.34 (5H, m), 7.63 (1H, d, J=15.76 Hz),7.66 (1H, d, J=7.80 Hz), 7.82 (1H, d, 7.80 Hz), 7.92 (1H, s). MS (m/z):393 [MH]⁺.

Example 48 Preparation ofN-Hydroxy-3-[2-(benzylamino-methyl)-ethyl-1H-benzimidazol-5-yl]-acrylamide(64)

The titled compound (64) was prepared according to the proceduresdescribed in Example 46, by using appropriate starting materials. HPLC:98.5%; t_(R)=(LC/PDA: Phenomenex Luna C18 2.0×150 mm 5μ column; 0.8mL/min, gradient 5-65% B over 15.5 min, Solvent A: H₂O with 0.1%trifluoroacetic acid; Solvent B: Acetonitrile with 0.1% trifluoroaceticacid; UV 254): 3.52 min. ¹H NMR (400 MHz, DMSO-d₆) δ 1.31 (3H, t) 3.37(2H, m), 3.50 (2H, t), 4.28 (4H, m), 6.48 (1H, d), 7.43-50 (3H, m), 7.55(3H, m) 7.73-7.83 (2H, t) 7.95 (1H, s), 9.25 (1H, bs), 10.76 (1H, bs).MS (m/z): 351 [MH]⁺.

Example 49 Preparation ofN-Hydroxy-3-(2-(benzyl-1-methyl-3-oxo-1H-benzimidazol-5-yl)-acrylamide

The titled compound (65) was prepared according to the proceduresdescribed in Example 42, by using appropriate starting materials. HPLC:99%; t_(R)=(LC/PDA: Phenomenex Luna C18 2.0×150 mm 5μ column; 0.8mL/min, gradient 5-65% B over 15.5 min, Solvent A: H₂O with 0.1%trifluoroacetic acid; Solvent B: Acetonitrile with 0.1% trifluoroaceticacid; UV 254): 4.48 min. ¹H NMR (400 MHz, DMSO-d₆) δ 3.87 (3H, s), 4.59(2H, s), 6.57 (1H, d, J=15.9 Hz), 7.09-7.36 (5H, m), 7.62 (1H, d, J=15.8Hz), 7.73-7.95 (3H, m). MS (m/z): 309 [MH]⁺.

Example 50 Preparation ofN-Hydroxy-3-[1-(2-diethylamino-ethyl)-2-phenethyl-1H-benzimidazol-5-yl]-acrylamide(66)

The titled compound (66) was prepared according to the proceduresdescribed in Example 1, by using appropriate starting materials. HPLC:100%; t_(R)=(LC/PDA: Phenomenex Luna C18 2.0×150 mm 5μ column; 0.8mL/min, gradient 5-65% B over 15.5 min, Solvent A: H₂O with 0.1%trifluoroacetic acid; Solvent B: Acetonitrile with 0.1% trifluoroaceticacid; UV 254): 3.72 min. ¹H NMR (400 MHz, CD₃OD) δ 1.29 (6H, t, J=7.3Hz), 3.26 (8H, m), 3.40 (2H, t, J=7.5 Hz), 4.60 (2H, t, J=8.0 Hz), 6.50(1H, d, J=16.0 Hz), 7.21 (5H, m), 7.62 (1H, d, J=16.0 Hz), 7.70 (2H, m),7.85 (1H, s); ¹³C NMR (100 MHz, CD₃OD) 39.0, 29.4, 34.3, 39.9, 48.4,50.3, 112.7, 116.6, 119.3, 125.8, 128.1, 129.6, 130.0, 133.9, 134.9,137.6, 140.8, 157.0, 166.0. MS (m/z): 407 [MH]⁺.

Example 51 Preparation ofN-Hydroxy-3-[2-phenethyl-1-(piperidin-1-yl-ethyl)-1H-benzimidazol-5-yl]-acrylamide(67)

The titled compound (67) was prepared according to the proceduresdescribed in Example 1, by using appropriate starting materials. HPLC:100%; t_(R)=(LC/PDA: Phenomenex Luna C18 2.0×150 mm 5μ column; 0.8mL/min, gradient 5-65% B over 15.5 min, Solvent A: H₂O O with 0.1%trifluoroacetic acid; Solvent B: Acetonitrile with 0.1% trifluoroaceticacid; UV 254): 3.90 min. ¹H NMR (400 MHz, CD₃OD) δ 1.86 (6H, br s), 3.26(8H, m), 3.40 (2H, t, J=7.5 Hz), 4.62 (2H, t, J=7.9 Hz), 6.50 (1H, d,J=16.0 Hz), 7.23 (5H, m), 7.62 (1H, d, J=16.0 Hz), 7.70 (2H), 7.84 (1H,s); ¹³C NMR (100 MHz, CD₃OD) δ 22.5, 24.2, 29.4, 34.3, 39.6, 54.4, 54.9,112.7, 116.6, 119.2, 125.7, 128.1, 129.6, 130.0, 133.8, 134.9, 137.8,140.8, 157.0, 166.0. MS (m/z): 419 [MH]⁺.

Example 52 Preparation ofN-Hydroxy-3-[2-phenyethyl-1-(2-pyrrolidin-1-yl-ethyl)-1H-benzimidazol-5-yl]-acrylamide(72)

The titled compound (72) was prepared according to the proceduresdescribed in Example 1, by using appropriate starting materials. HPLC:100%; t_(R)=(LC/PDA: Phenomenex Luna C18 2.0×150 mm 5μ column; 0.8mL/min, gradient 5-65% B over 15.5 min, Solvent A: H₂O with 0.1%trifluoroacetic acid; Solvent B: Acetonitrile with 0.1% trifluoroaceticacid; UV 254): 3.71 min. ¹H NMR (400 MHz, CD₃OD) δ 2.06 (4H, br), 3.21(2H, t, J=7.4 Hz), 3.26 (4H, m), 3.37 (2H, t, J=7.7 Hz), 3.42 (2H, t,J=7.5 Hz), 4.57 (2H, t, J=7.4 Hz), 6.47 (1H, d, J=16.0 Hz), 7.21 (5H,m), 7.58 (1H, d, J=16.0 Hz), 7.67 (1H, d, J=8.6 Hz), 7.74 (1H, d, J=8.6Hz), 7.83 (1H, s); ¹³C NMR (100 MHz, CD₃OD) δ 24.1, 29.4, 34.3, 41.1,52.8, 55.7, 112.9, 116.5, 119.2, 125.8, 128.1, 129.6, 130.0, 133.9,134.9, 137.2, 140.7, 140.8, 157.0, 165.9. MS (m/z): 405 [MH]⁺.

Example 53 Preparation ofN-Hydroxy-3-[2-(2-benzylamino-ethyl)-1-ethyl-1H-benzimidazol-5-yl]-acrylamide(74)

The titled compound (74) was prepared according to the proceduresdescribed in Example 46, by using appropriate starting materials. HPLC:98.5%; t_(R)=(LC/PDA: Phenomenex Luna C18 2.0×150 mm 5μ column; 0.8mL/min, gradient 5-65% B over 15.5 min, Solvent A: H₂O with 0.1%trifluoroacetic acid; Solvent B: Acetonitrile with 0.1% trifluoroaceticacid; UV 254): 3.52 min. ¹H NMR (400 MHz, DMSO-d₆) δ 1.31 (3H, t) 3.37(2H, m), 3.50 (2H, t), 4.28 (4H, m), 6.48 (1H, d), 7.43-50 (3H, m), 7.55(3H, m) 7.73-7.83 (2H, t) 7.95 (1H, s), 9.25 (1H, bs), 10.76 (1H, bs).MS (m/z): 365 [MH]⁺.

Example 54 Preparation ofN-Hydroxy-3-[2-phenethyl-1-(3-pyrrolidin-1-yl-propyl)-1H-benzimidazol-5-yl]-acrylamide(82)

The titled compound (82) was prepared according to the proceduresdescribed in Example 1, by using appropriate starting materials. HPLC:100%; t_(R)=(LC/PDA: Phenomenex Luna C18 2.0×150 mm 5μ column; 0.8mL/min, gradient 5-65% B over 15.5 min, Solvent A: H₂O with 0.1%trifluoroacetic acid; Solvent B: Acetonitrile with 0.1% trifluoroaceticacid; UV 254): 1.18 min. ¹H NMR (400 MHz, CD₃OD) δ 2.01 (2H), 2.17 (4H),3.03 (2H), 3.26 (4H), 3.48 (2H), 3.62 (2H), 4.37 (2H), 6.60 (1H), 7.27(5H), 7.71 (1H), 7.78 (2H), 7.91 (1H). MS (m/z): 419 [MH]⁺.

Example 55 Preparation ofN-Hydroxy-3-[1-(3-Dimethylamino-2,2-dimethyl-propyl)-2-(2-pyridin-3-yl-ethyl)-1H-benzimidazol-5-yl]-acrylamide(86)

The titled compound (86) was prepared according to the proceduresdescribed in Example 42, by using appropriate starting materials. HPLC:90.4%; t_(R)=(LC/PDA: Phenomenex Luna C18 2.0×150 mm 5μ column; 0.8mL/min, gradient 5-65% B over 15.5 min, Solvent A: H₂O with 0.1%trifluoroacetic acid; Solvent B: Acetonitrile with 0.1% trifluoroaceticacid; UV 254): 1.24 min. ¹H NMR (400 MHz, DMSO-d₆) δ 1.00 (6H, s), 2.94(6H, s), 3.32 (2H, m), 3.38 (4H, m) 4.35 (2H, m), 6.52 (1H, d),7.58-7.86 (5H, m) 8.20 (1H, d), 8.65 (1H, m) 8.77 (1H, s), 9.50 (1H, s).MS (m/z): 422 [MH]⁺.

Example 56 Preparation of2-[2-Phenethyl-1-(3,4,5-trimethoxy-benzyl)-1H-benzimidazol-5-yl]-cyclopropanecarboxylicacid hydroxyamide (88) Step 1

To a solution of (CH₃)₃S(O)I (132 mg, 0.6 mmol) in anhydrous DMSO (1 mL)was added sodium hydride (28 mg, 60% in mineral oil) at room temperatureunder nitrogen gas, then a solution of the compound (244 mg, 0.5 mmol),3-[2-phenethyl-1-(3,4,5-trimethoxy-benzyl)-1H-benzimidazol-5-yl]-acrylicacid methyl ester (prepared according to Example 1, step 1-3), in 4 mLof anhydrous THF was added after 10 mins. The resulting mixture was thenstirred at room temperature overnight. After an aqueous work-up, theresidue was obtained as oil (135 mg), which was then subjected to nextstep without further purification.

Step 2

To a solution of above crude product in 0.5 mL MeOH was added apre-prepared 2.0 M NH₂OH stock solution as we did before (2 mL). Theresulting mixture was stirred at room temperature for 4 hrs. Afterquenching with TFA (0.4 mL), the resulting mixture was subjected to HPLCpurification to afford 10 mg of desired titled compound (88). HPLC: 99%;t_(R)=(LC/PDA: Phenomenex Luna C18 2.0×150 mm 5μ column; 0.8 ml/min,gradient 5-65% B over 15.5 min, Solvent A: H₂O with 0.1% trifluoroaceticacid; Solvent B: Acetonitrile with 0.1% trifluoroacetic acid; UV 254):6.36 min. ¹H NMR (400 MHz, CD₃OD) δ 1.21-1.29 (1H, m), 1.45-1.52 (1H,m), 1.75-1.79 (1H, m), 2.48-2.55 (1H, m), 2.99 (2H, t, J=8.0 Hz), 3.45(2H, t, J=8.0 Hz), 3.61 (6H, s), 3.64 (3H, s), 5.42 (2H, s), 6.40 (2H,s), 7.00-7.18 (5H, m), 7.26 (1H, d, J=8.4 Hz), 7.45 (1H, s), 7.59 (1H,d, J=8.4 Hz). MS (m/z): 502 [MH]⁺.

Example 57 Preparation ofN-Hydroxy-3-[2-benzylsulfanyl-1-(3-dimethylamino-2,2-dimethyl-propyl)-1H-benzimidazol-5-yl]-acrylamide(89) Step 1

3-[4-(3-dimethylamino-2,2-dimethyl-propylamino)-3-nitro-phenyl]-acrylicacid (1.93 g, 6.0 mmol, prepared as described in Example 1, step 1), Tinchloride (13.5 g, 60 mmol) and MeOH (50 mL) was mixed and heated at 45°C. for 20 hours. The reaction mixture was cooled to room temperature andsolvent was removed under reduced pressure. To the residue was added 100mL dichloromethane and 100 mL water. The pH was adjusted to withconcentrated ammonia. The layers were separated, and the aqueous phasewas extracted with 100 mL dichloromethane. The organic extracts werecombined, dried over sodium sulfate, filtered and the solvent wasremoved under reduced pressure. To the resulting residue was added MeOH(100 mL), CS₂ (18 mL) and potassium hydroxide (3.4 g. The reactionmixture was heated at 80° C. for 16 hours, then cooled to roomtemperature and the solvents were removed under reduced pressure. Theresulting crude product was recrystallized from MeOH.

The product,3-[1-(3-dimethylamino-2,2-dimethyl-propyl)-2-thioxo-2,3-dihydro-1H-benzimidazol-5-yl]-acrylicacid, was obtained in 75% yield in two steps (1.5 g). MS (m/z): 334[MH]⁺.

Step 2

3-[1-(3-dimethylamino-2,2-dimethyl-propyl)-2-thioxo-2,3-dihydro-1H-benzimidazol-5-yl]-acrylicacid (100 mg, 0.3 mmol), benzyl bromide (360 mg, 3.6 mmol), andpotassium carbonate (0.83 g) were mixed with 10 mL DMF. The resultingmixture was stirred overnight at 45° C. The desired product,3-[2-benzylsulfanyl-1-(3-dimethylamino-2,2-dimethyl-propyl)-2,3-dihydro-1H-benzimidazol-5-yl]-acrylicacid benzyl ester, was purified by preparative HPLC: 150 mg (yield,76.6%). ¹H NMR (400 MHz, DMSO-d₆) δ 1.08 (6H, s), 2.88 (3H, s), 2.89(3H, s), 3.30 (2H), 4.11 (2H, s), 4.65 (2H, s), 5.24 (2H, s), 6.72 (2H,d, J=15.96 Hz), 7.26-7.47 (10H, m), 7.68 (2H, bs), 7.83 (1H, d, J=15.96Hz), 8.00 (1H, s). MS (m/z): 514 [MH]⁺.

Step 3

The titled compound (89) was obtained by treating3-[2-benzylsulfanyl-1-(3-dimethylamino-2,2-dimethyl-propyl)-2,3-dihydro-1H-benzimidazol-5-yl]-acrylicacid benzyl ester according to method previously described for thepreparation of hydroxamic acid (Step 4 of Example 1). HPLC: 99%;t_(R)=(LC/PDA: Phenomenex Luna C18 2.0×150 mm 5μ column; 0.8 mL/min,gradient 5-65% B over 15.5 min, Solvent A: H₂O with 0.1% trifluoroaceticacid; Solvent B: Acetonitrile with 0.1% trifluoroacetic acid; UV 254):2.87 min. ¹H NMR (400 MHz, DMSO-d₆) δ 1.09 (6H, s), 2.88 (3H, s), 2.89(3H, s), 3.26 (2H), 4.11 (2H, s), 4.65 (2H, s), 6.48 (2H, d, J=15.79Hz), 7.26-7.47 (6H, m), 7.58 (1H, d, J=15.79 Hz), 7.65 (1H, d, J=8.48Hz), 7.80 (1H, s). MS (m/z): 439 [MH]⁺.

Example 58 Preparation ofN-Hydroxy-3-[1-(3-dimethylamino-2,2-dimethyl-propyl)-2-phenylmethanesulfonyl-1H-benzimidazol-5-yl]-acrylamide(91) Step 1

118 mg of3-[2-benzylsulfanyl-1-(3-dimethylamino-2,2-dimethyl-propyl)-2,3-dihydro-1H-benzimidazol-5-yl]-acrylicacid benzyl ester (prepared according Example 57, step 1-2), 1.0 mL ofhydrogen peroxide (30%) and 10 mL of acetic acid were mixed at 0° C. inan ice bath. Without adding additional ice, the reaction mixture wasstirred overnight. The product,3-[1-(3-dimethylamino-2,2-dimethyl-propyl)-2-phenylmethanesulfinyl-2,3-hydro-1H-benzimidazol-5-yl]-acrylicacid benzyl ester, was obtained quantitatively. MS (m/z): 530 [MH]⁺.

Step 2

The titled compound (91) was obtained by treating3-[1-(3-dimethylamino-2,2-dimethyl-propyl)-2-phenylmethanesulfinyl-2,3-hydro-1H-benzimidazol-5-yl]-acrylicacid benzyl ester according to the method previously described for thepreparation of hydroxamic acid (Step 4 of Example 1). HPLC: 77.1%;t_(R)=(LC/PDA: Phenomenex Luna C18 2.0×150 mm 5μ column; 0.8 mL/min,gradient 5-65% B over 15.5 min, Solvent A: H₂O with 0.1% trifluoroaceticacid; Solvent B: Acetonitrile with 0.1% trifluoroacetic acid; UV 254):1.46 min. ¹H NMR (400 MHz, DMSO-d₆) δ 1.11 (6H, s), 2.90 (3H, s), 2.91(3H, s), 3.12 (2H, s), 3.82 (2H, s), 4.79 (2H, s), 6.56 (1H, d, J=15.80Hz), 7.15-7.32 (5H, m), 7.59-7.66 (2H, m), 7.87 (1H, d, J=8.68 Hz), 8.06(1H, s). MS (m/z): 455 [MH]⁺.

Example 59 Preparation ofN-Hydroxy-3-(2-benzyl-1-ethyl-1H-benzimidazol-5-yl)-acrylamide (92)

The titled compound (92) was prepared according to the proceduresdescribed in Example 42, by using appropriate starting materials. HPLC:97.0%; t_(R)=(LC/PDA: Phenomenex Luna C18 2.0×150 mm 5μ column; 0.8ml/min, gradient 5-65% B over 15.5 min, Solvent A: H₂O with 0.1%trifluoroacetic acid; Solvent B: Acetonitrile with 0.1% trifluoroaceticacid; UV 254): 1.60 min. ¹H NMR (400 MHz, DMSO-d₆) δ 1.17 (3H, t, J=7.1Hz), 4.34 (2H, dd, J=6.8 Hz), 4.56 (2H, s), 6.55 (1H, d, J=15.8 Hz),7.31-7.40 (5H, m), 7.63 (1H, d, J=15.8 Hz), 7.85-7.93 (3H, m). MS (m/z):322 [MH]⁺.

Example 60 Preparation ofN-Hydroxy-3-(1-ethyl-2-[3-(1H-indol-3-yl)-propyl]-1H-benzimidazol-5-yl)-acrylamide(93)

The titled compound (93) was prepared according to the proceduresdescribed in Example 42, by using appropriate starting materials. HPLC:98.5%; t_(R)=(LC/PDA: Phenomenex Luna C18 2.0×150 mm 5μ column; 0.8mL/min, gradient 5-65% B over 15.5 min, Solvent A: H₂O with 0.1%trifluoroacetic acid; Solvent B: Acetonitrile with 0.1% trifluoroaceticacid; UV 254): 1.98 min. ¹H NMR (400 MHz, DMSO-d₆) δ 1.33 (3H, t), 2.22(2H, m), 2.87 (2H, t), 3.16 (2H, m), 4.37 (2H, m), 6.53 (1H, d), 6.98(1H, m) 7.06 (1H, m) 7.19 (1H, s), 7.33 (1H, d), 7.54-7.88 (5H, d),10.82 (2H, bs). MS (m/z): 389 [MH]⁺.

Example 61 Preparation ofN-Hydroxy-3-(1-(3-dimethylamino-2,2-dimethyl-propyl)-2-[2-(3-methoxy-phenyl)-ethyl]-1H-benzimidazol-5-yl)-acrylamide(94)

The titled compound (94) was prepared according to the proceduresdescribed in Example 42, by using appropriate starting materials. HPLC:99.7%; t_(R)=(LC/PDA: Phenomenex Luna C18 2.0×150 mm 5μ column; 0.8mL/min, gradient 5-65% B over 15.5 min, Solvent A: H₂O with 0.1%trifluoroacetic acid; Solvent B: Acetonitrile with 0.1% trifluoroaceticacid; UV 254): 1.34 min. ¹H NMR (400 MHz, DMSO-d₆) δ 1.03 (6H, s), 2.90(6H, s), 3.19 (2H, t), 3.34 (4H, s) 3.71 (3H, s) 4.29 (2H, t), 6.52 (1H,d), 6.80 (1H, m) 6.88 (2H, d) 7.22 (1H, m), 7.62 (2H, m), 7.83-7.89 (2H,m), 9.34 (1H, s), 10.77 (1H, bs). MS (m/z): 451 [MH]⁺.

Example 62 Preparation ofN-Hydroxy-3-[1-ethyl-2-(3-phenoxy-propyl)-1H-benzimidazol-5-yl]-acrylamide(96)

The titled compound (96) was prepared according to the proceduresdescribed in Example 46, by using appropriate starting materials. HPLC:99.6%; t_(R)=(LC/PDA: Phenomenex Luna C18 2.0×150 mm 5μ column; 0.8mL/min, gradient 5-65% B over 15.5 min, Solvent A: H₂O with 0.1%trifluoroacetic acid; Solvent B: Acetonitrile with 0.1% trifluoroaceticacid; UV 254): 1.83 min, ¹H NMR (400 MHz, DMSO-d₆) δ 1.36 (3H, t), 2.32(2H, m), 3.34 (2H, m), 4.12 (2H, m), 4.46 (2H, m), 6.58 (1H, d), 6.73(2H, d) 6.90 (1H, m) 7.22 (2H, m), 7.65 (1H, d), 7.80 (1H, d), 7.94 (2H,m). MS (m/z): 366 [MH]⁺.

Example 63 Preparation ofN-Hydroxy-3-(2-{[2-(4-methoxy-phenyl)-acetylamino]-methyl}-1-methyl-H-benzimidazol-5-yl)-acrylamide(99)

The titled compound (99) was prepared according to the proceduresdescribed in Example 42, by using appropriate starting materials. HPLC:97.0%; t_(R)=(LC/PDA: Phenomenex Luna C18 2.0×150 mm 5μ column; 0.8mL/min, gradient 5-65% B over 15.5 min, Solvent A: H₂O with 0.1%trifluoroacetic acid; Solvent B: Acetonitrile with 0.1% trifluoroaceticacid; UV 254): 2.75 min. ¹H NMR (400 MHz, DMSO-d₆) δ 3.48 (2H, s), 3.67(3H, s), 3.87 (3H, s), 4.71 (2H, m), 6.55 (1H, d), 6.86 (3H, m) 7.18(3H, m) 7.84-7.92 (2H, m), 10.77 (1H, s). MS (m/z): 395 [MH]⁺.

Example 64 Preparation of2-(1-Methyl-2-phenethyl-1H-benzimidazol-5-yl)-cyclopropanecarboxylicacid hydroxyamide (100)

The titled compound (100) was prepared according to the proceduresdescribed in Example 56, by using appropriate starting materials. HPLC:99%; t_(R)=(LC/PDA: Phenomenex Luna C18 2.0×150 mm 5μ column; 0.8mL/min, gradient 5-65% B over 15.5 min, Solvent A: H₂O with 0.1%trifluoroacetic acid; Solvent B: Acetonitrile with 0.1% trifluoroaceticacid; UV 254): 6.36 min ¹H NMR (400 MHz, CDCl₃, with one drop ofd₆-DMSO-d₆) δ 1.25 (1H, m), 1.64 (1H, m), 1.88 (1H, m), 1.98 (3H, s),2.63 (1H, m), 3.23 (2H, t, J=8.0 Hz), 3.52 (2H, t, J=8.0 Hz), 7.08-7.45(7H, m), 7.57 (1H, s). MS (m/z): 336 [MH]⁺.

Example 65 Preparation ofN-Hydroxy-3-(1-methyl-1H-benzimidazol-5-yl)-acrylamide (49)

The titled compound (49) was prepared according to the proceduresdescribed in Example 1, by using appropriate starting materials. HPLC:99%; t_(R)=(LC/PDA: Phenomenex Luna C18 2.0×150 mm 5μ column; 0.8mL/min, gradient 5-65% B over 15.5 min, Solvent A: H₂O with 0.1%trifluoroacetic acid; Solvent B: Acetonitrile with 0.1% trifluoroaceticacid; UV 254): 1.05 min. ¹H NMR: (400 MHz, CD₃OD) δ 4.05 (3H, s), 6.52(1H, d, J=15.8 Hz), 7.62 (1H, d, J=15.8 Hz), 7.77-7.89 (3H, m), 9.19(1H, s). MS (m/z): 218 [MH]⁺.

The following compounds are some representative examples prepared bymethods disclosed or analogous to those disclosed in above Examples1-65:

TABLE 1 m/z Compound Structures [MH]⁺ 1

380 2

488 3

431 4

474 5

354 6

383 7

490 8

382 9

355 10

325 11

339 12

399 13

339 14

366 15

380 16

399 17

421 18

413 19

382 20

344 21

318 22

365 23

374 24

344 25

364 26

412 27

413 28

429 29

320 30

433 31

435 32

380 33

398 34

350 35

294 36

350 37

246 38

336 39

232 40

340 41

427 42

309 43

246 44

421 45

490 46

304 47

262 48

322 49

218 50

308 51

204 52

336 53

232 54

365 55

352 56

416 57

407 58

398 59

322 60

337 61

427 62

367 63

393 64

351 65

309 66

407 67

419 68

441 69

383 70

393 71

415 72

405 73

409 74

365 75

378 76

481 77

352 78

394 79

348 80

408 81

448 82

419 83

435 84

439 85

326 86

422 87

309 88

502 89

439 90

315 91

455 92

322 93

389 94

451 95

338 96

366 97

336 98

297 99

395 100

336

By methods analogous to those disclosed above, a wide variety ofcompounds of Formula I could be prepared, including, but not limited to,those in Table 2 (a):

TABLE 2 (a) 101

N-Hydroxy-3-[1-methyl-2-(2-piperidin- 1-yl-ethyl)-1H-benzimidazol-5-yl)-acrylamide 102

N-Hydroxy-3-[2-(2-diethylamino- ethyl)-1-methyl-1H-benzimidazol-5-yl]-acrylamide 103

N-Hydroxy-3-[2-(2-cyclohexyl-ethyl)- 1-(2-pyridin-1-yl-ethyl)-1H-benzimidazol-5-yl]-acrylamide 104

N-Hydroxy-N-methyl-3-[2-(2- cyclohexyl-ethyl)-1-(2-pyrrolidin-1-yl-ethyl)-1H-benzimidazol-5-yl]- acrylamide 105

N-Hydroxy-[2-(2-cyclohexyl-ethyl)-1- methyl-1H-benzimidazol-5-yl]-acrylamide 106

N-Hydroxy-[2-(2-cyclopenthyl-ethyl)-1- (2-pyrrolidin-1-yl-ethyl)-1H-benzimidazol-5-yl]-acrylamide 107

N-Hydroxy-3-[1-methyl-2-(2- pyrrolidin-1-yl-ethyl)-1H-benzimidazol-5-yl]-acrylamide 108

(L)-N-Hydroxy-3-[2-(1-benzylamino-2- phenyl-ethyl)-1-methyl-1H-benzimidazol-5-yl]-acrylamide 109

N-Hydroxy-3-[2-benzyloxy-1-(2- pyrrolidin-1-yl)-1H-benzimidazol-5-yl]-acrylamide 110

N-Hydroxy-3-(2-benzylsulfanyl-1- methyl-1H-benzimidazol-5-yl)-acrylamide 111

N-Hydroxy-3-[2-phenethylamino-1-(2- pyrrolidin-1-yl-ethyl)-1H-benzimidazol-5-yl]-acrylamide 112

N-Hydroxy-3-(1-methyl-2-quinolin-3- ylmethyl-1H-benzimidazol-5-yl)-acrylamide 113

N-Hydroxy-3-[1-(2-pyrrolidin-1-yl- ethyl)-2-(2-thiophen-2-yl-ethyl)-1H-benzimidazol-5-yl]-acrylamide 114

N-Hydroxy-3-[1-methyl-2-(2- naphthalen-2-yl-ethyl)-1H-benzimidazol-5-yl]-acrylamide 115

N-Hydroxy-3-(4,7-Dimethyl-2- phenethyl-1-pyridin-2-ylmethyl-1H-benzimidazol-5-yl)-acrylamide 116

N-Hydroxy-3-(7-benzyloxy-4-methyl- 2-phenethyl-1-phenethyl-1-pyridin-2ylmethyl-1H-benzimidazol-5-yl)- acrylamide 117

N-Hydroxy-3-(4,7-difluoro-2- phenethyl-1-pyridin-2ylmethyl-1H-benzimidazol-5-yl)-acrylamide

By methods analogous to those disclosed above and by varying thestarting materials used in the synthesis, a wide variety of compounds ofFormula I could be prepared, including, but not limited to, those inTable 2 (b):

TABLE 2(b) R₁ R₂ R₃ R₄ X Y 118 H

H H H CH₃ 119 H

CH₃ H F OCH₃ 120 H

H CH₃ Cl CH₃ 121 H

CH₃ CH₃ Br H 122 H

H H CH₃ F 123 H

CH₃ H OCH₃ Cl 124 Propyl

H CH₃ CF₃ Br 125 Propyl

CH₃ CH₃ CN CH₃ 126 Propyl

H H OCF₃ OCH₃ 127 Propyl

CH₃ H NO₂ CF₃ 128 Propyl

H CH₃ CH₃ CN 129 Propyl

CH₃ CH₃ OCH₃ OCF₃ 130 Propyl

H H F NO₂ 131 Propyl

CH₃ H CH₃ CH₃ 132

H CH₃ OCH₃ OCH₃ 133

CH₃ CH₃ F H 134

H H CH₃ F 135

CH₃ H OCH₃ Cl 136

H CH₃ F Br 137

CH₃ CH₃ CH₃ CH₃ 138

H H CH₃ OCH₃ 139

CH₃ H OCH₃ CF₃ 140

H CH₃ F CN 141

CH₃ CH₃ F OCF₃ 142

H H Cl NO₂ 143

CH₃ H Br F 144

H CH₃ CH₃ CH₃ 145

CH₃ CH₃ OCH₃ OCH₃ 146

H H CF₃ CH₃ 147

CH₃ H CN H 148

H CH₃ OCF₃ F 149

CH₃ CH₃ NO₂ Cl 150

H H CH₃ Br 151

CH₃ H CH₃ CH₃ 152

H CH₃ OCH₃ OCH₃ 153

CH₃ CH₃ F CF₃ 154

H H CH₃ CN 155

CH₃ H OCH₃ OCF₃ 156

H CH₃ CH₃ NO₂ 157

CH₃ CH₃ OCH₃ CH₃ 158

H H F F 159

CH₃ H H CH₃ 160

H CH₃ F OCH₃ 161

CH₃ CH₃ Cl CH₃ 162

H H Br H 163

CH₃ H CH₃ F 164

H CH₃ OCH₃ Cl 165

CH₃ CH₃ CF₃ Br 166

H H CN CH₃ 167

CH₃ H OCF₃ OCH₃ 168

H CH₃ NO₂ CF₃ 169

CH₃ CH₃ CH₃ CN 170

H H OCH₃ OCF₃ 171

CH₃ H F NO₂ 172

H CH₃ CH₃ CH₃ 173

CH₃ CH₃ OCH₃ H 174

H H H F 175

CH₃ H F Cl 176

H CH₃ Cl Br 177

CH₃ CH₃ Br CH₃ 178

H H CH₃ OCH₃ 179

CH₃ H OCH₃ CF₃ 180

H CH₃ CF₃ CN 181

CH₃ CH₃ CN OCF₃ 182

H H OCF₃ NO₂ 183

CH₃ H NO₂ F 184

H CH₃ CH₃ CH₃ 185

CH₃ CH₃ OCH₃ OCH₃ 186

H H F H 187

CH₃ H CH₃ F 188

H CH₃ OCH₃ Cl 189

CH₃ CH₃ CH₃ Br 190

H H OCH₃ CH₃ 191

CH₃ H CH₃ OCH₃ 192

H CH₃ CH₃ CF₃ 193

CH₃ CH₃ OCH₃ CN 194

H H F OCF₃ 195

CH₃ H H NO₂ 196

H CH₃ F CH₃ 197

CH₃ CH₃ Cl OCH₃ 198

H H Br F 199

CH₃ H CH₃ CH₃ 200

H CH₃ OCH₃ OCH₃ 201

CH₃ CH₃ CF₃ CH₃ 202

H H CN F 203

CH₃ H OCF₃ CH₃ 204

H CH₃ NO₂ OCH₃ 205

CH₃ CH₃ CH₃ CH₃ 206

H H OCH₃ H 207

CH₃ H CH₃ F 208

H CH₃ OCH₃ Cl 209

CH₃ CH₃ F Br 210

H H CH₃ CH₃ 211

CH₃ H H OCH₃ 212

H CH₃ F CF₃ 213

CH₃ CH₃ Cl CN 214

H H Br OCF₃ 215

CH₃ H CH₃ NO₂ 216

H H OCH₃ CH₃ 217

CH₃ H CF₃ F 218

H CH₃ CN CH₃ 219

CH₃ CH₃ OCF₃ OCH₃ 220

H H NO₂ CH₃ 221

CH₃ H CH₃ OCH₃ 222

H CH₃ H H 223

CH₃ CH₃ F F 224

H H Cl Cl 225

CH₃ H Br Br 226

H CH₃ CH₃ CH₃ 227

CH₃ CH₃ OCH₃ OCH₃ 228

H H CF₃ CF₃ 229

CH₃ H CN CN 230

H CH₃ OCF₃ OCF₃ 231

CH₃ CH₃ NO₂ NO₂ 232

H H CH₃ CH₃ 233

CH₃ H OCH₃ OCH₃ 234

H CH₃ F F 235

CH₃ CH₃ CH₃ CH₃ 236

H H OCH₃ OCH₃ 237

CH₃ H CH₃ F 238

H CH₃ OCH₃ CH₃ 239

CH₃ CH₃ F OCH₃

Biological Testing and Enzyme Assays Recombinant GST-HDAC1 ProteinExpression and Purification

Human cDNA library was prepared using cultured SW620 cells.Amplification of human HDAC1 and HDAC8 coding region from this cDNAlibrary was cloned separately into the baculovirus expression pDEST20vector and pFASTBAC vector respectively (GATEWAY Cloning Technology,Invitrogen Pte Ltd). The pDEST20-HDAC1 and pFASTBAC-HTGST-HDAC8constructs were confirmed by DNA sequencing. Recombinant baculovirus wasprepared using the Bac-To-Bac method following the manufacturer'sinstruction (Invitrogen Pte Ltd). Baculovirus titer was determined byplaque assay to be about 10⁸ PFU/ml.

Expression of GST-HDAC1 or HTGST-HDAC8 was done by infecting SF9 cells(Invitrogen Pte Ltd) with pDEST20-HDAC1 or pFASTBAC-GST-HDAC8baculovirus at MOI=1 for 48 h. Soluble cell lysate was incubated withpre-equilibrated Glutathione Sepharose 4B beads (Amersham) at 4° C. for2 h. The beads were washed with PBS buffer for 3 times. The GST-HDAC1protein or GST-HDAC8 protein was eluted by elution buffer containing 50mM Tris, pH8.0, 150 mM NaCl, 1% Triton X-100 and 10 mM or 20 mM reducedGlutathione. The purified GST-HDAC1 protein or purified GST-HDAC8protein was dialyzed with HDAC storage buffer containing 10 mM Tris,pH7.5, 100 mM NaCl and 3 mM MgCl₂. 20% Glycerol was added to purifiedGST-HDAC1 protein or purified GST-HDAC8 before storage at −80° C.

In Vitro HDAC Assay for Determination of IC50 Values

The assay has been carried out in 96 well format and the BIOMOLfluorescent-based HDAC activity assay has been applied. The reactioncomposed of assay buffer, containing 25 mM Tris pH 7.5, 137 mM NaCl, 2.7mM KCl, 1 mM MgCl₂, 1 mg/ml BSA, tested compounds, 500 nM HDAC8 enzymeor 600 nM HDAC1 enzyme, 200 μM Flur de lys p53 peptide substrate forHDAC8 enzyme or 500 μM Flur de lys generic substrate for HDAC1 enzymeand subsequently was incubated at room temperature for 2 h. Flur de lysDeveloper was added and the reaction was incubated for 10 min. Briefly,deacetylation of the substrate sensitizes it to the developer, whichthen generates a fluorophore (symbol). The fluorophore is excited with360 nm light and the emitted light (460 nm) is detected on afluorometric plate reader (Tecan Ultra Microplate detection system,Tecan Group Ltd.).

The analytical software, Prism 3.0 has been used to generate IC₅₀ from aseries of data. The HDAC enzyme inhibition results of representativecompounds are shown in Table 3.

TABLE 3 HDAC1 Enzyme HDAC8 Enzyme Activity Activity Compound IC₅₀ (μM)IC₅₀ (μM) 1 0.051 0.119 2 0.026 0.355 3 1.37 1.71 4 1.34 0.790 5 4.320.401 6 1.38 0.262 7 1.52 0.336 8 0.286 0.454 9 1.34 0.344 10 2.66 0.88311 0.846 0.161 12 0.131 0.202 13 0.385 0.141 14 0.171 0.251 15 0.2060.313 16 0.194 0.366 17 0.024 0.353 18 0.438 0.290 19 0.165 0.145 201.91 0.537 21 0.064 0.238 22 1.326 0.234 23 0.529 0.402 24 3.24 0.203 251.32 0.601 26 0.876 1.005 27 0.092 0.329 28 0.206 0.300 29 49.06 33.9630 0.195 0.724 31 0.246 1.09 32 2.21 1.89 33 0.449 1.45 34 1.46 0.846 350.371 0.412 36 0.227 37 0.897 38 0.218 0.148 39 1.22 0.201 40 3.30 0.44141 0.195 0.159 42 0.479 0.237 43 0.947 0.192 44 0.268 0.345 45 0.167 461.67 47 1.09 48 0.356 0.291 49 1.40 50 0.173 51 0.896 52 0.160 53 1.8554 0.100 55 0.137 56 0.158 57 0.153 58 1.14 59 0.382 60 0.116 61 0.19662 0.234 63 0.162 64 0.230 65 0.062 66 0.072 0.255 67 0.039 0.254 680.294 69 0.146 70 0.923 71 0.167 72 0.052 73 0.560 74 0.371 75 0.290 761.03 77 0.570 78 >100 79 1.26 80 1.69 81 1.60 82 0.304 83 0.071 84 0.05485 0.131 86 0.400 87 0.517 88 0.297 89 0.116 90 0.166 91 0.030 92 0.16893 0.065 94 0.052 95 0.061 96 0.125

Cell-Based Proliferation Assay for Determination of GI₅₀ Values

Human colon cancer cell lines (Colo205 and HCT116), human breast cancercell lines (MDA-MB435 and MDA-MB231), and human lung cancer cell line(A549) were obtained from ATCC. Colo205 cells were cultivated in RPMI1640 containing 2 mM L-Glutamine, 5% FBS, 1.0 mM Na Pyruvate. A549 andMDA-MB231 were cultivated in RPMI 1640 containing 2 mM L-glutamine, 5%FBS. MDA-MB435 cells were cultivated in DMEM containing 2 mML-Glutamine, 5% FBS. HCT116 cells were cultivated in IMEM containing 2mM L-Glutamine, 5% FBS. A549 and Colo205 cells were seeded in 96-wellsplate at 2000 and 5000 cells per well respectively. MDA-MB435, HCT116,MDA-MB231 cells were seeded in 96-wells plate at 6000 cells per well.The plates were incubated at 37° C., 5% CO₂, for 24 h. Cells weretreated with compounds at various concentration for 96 h. Cell growthwas then monitored using cyquant cell proliferation assay (InvitrogenPte Ltd). Dose response curves were plotted to determine GI₅₀ values forthe compounds using XL-fit.

The cell activity results of representative compounds are shown in Table4. Table 5 summarized the antiproliferative activities of selectedcompounds including their different salts for additional cancer celllines. These data indicate that compounds in this invention are highlyactive in inhibition of tumor cell growth.

TABLE 4 GI50 GI50 Compound (Colo 205, μM) (MDA-MB435, μM) 1 0.52 1.64 20.43 0.32 4 29.87 25.70 5 >100 6 >100 7 >100 8 41.36 58.42 9 >100 >10011 >100 >100 12 0.38 1.07 13 12.32 14.05 14 3.07 5.99 15 1.99 4.07 160.94 0.98 17 0.06 0.56 18 4.69 6.16 19 4.10 3.97 20 30.86 37.22 21 25.9130.26 22 13.47 13.35 23 3.65 3.72 24 30.70 35.02 25 8.10 6.82 26 8.796.67 27 2.23 3.44 28 2.53 5.15 30 11.44 19.85 31 1.87 4.06 33 1.54 3.3835 1.89 6.76 36 2.29 2.17 37 7.82 7.90 38 1.47 1.53 39 11.68 12.05 4025.62 30.97 41 1.65 1.91 42 14.41 15.75 43 9.18 8.62 44 2.82 3.65 452.41 1.90 48 1.45 1.78 50 4.29 5.19 52 2.04 3.58 54 4.47 5.9255 >100 >100 56 >100 >100 57 1.11 1.39 59 2.72 3.69 60 2.47 3.60 61 2.693.05 62 11.65 19.80 63 2.00 64 1.70 65 36.89 66 0.22 67 0.08 68 0.73 697.16 70 2.90 71 7.09 72 0.18 73 6.67 74 2.07 75 2.88 82 0.72 83 0.25 840.17 85 1.65 86 13.13 87 47.71 88 1.26 89 0.12

TABLE 5 Compound 17 Compound 2 Salt of Compound 67 Free Salt of FreeSalt of Methane Salt of Free Salt of Activity base CF3COOH base HClsulfonic acid CF3COOH base CF3COOH IC₅₀ 0.043 0.049 0.029 0.044 0.0510.024 0.037 0.039 (HDAC1, μM) IC₅₀ 0.064 0.029 0.056 (HDAC3, μM) IC₅₀0.267 0.353 0.254 (HDAC8, μM) GI₅₀ 0.4 0.4 0.06 0.06 0.04 0.11 0.09 0.09(Colo205, μM) GI₅₀ 0.4 0.3 0.06 (HCT116, μM) GI₅₀ 0.3 0.6 0.19 (MDA-MB435, μM) GI₅₀ 0.5 0.7 0.06 (MDA- MB231, μM) GI₅₀ 0.3 0.2 0.08 (A549,μM)

Histone H3 Acetylation Assay

A hallmark of histone deacetylase (HDAC) inhibition is the increase inthe acetylation level of histones. Histone acetylation, including H3, H4and H2A can be detected by immuno-blotting (western-blot). Colo205cells, approximately 1.5×10⁶ cells/10 cm dish, were seeded in thepreviously described medium, cultivated for 24 h and subsequentlytreated with HDAC inhibitory agents at 0.1, 1, 5 and 10 μM finalconcentration. After 24 h, cells were harvested and lysed according tothe instruction from Sigma Mammalian Cell Lysis Kit. The proteinconcentration was quantified using BCA method (Sigma Pte Ltd). Theprotein lysate was separated using 4-12% bis-tris SDS-PAGE gel(Invitrogen Pte Ltd) and was transferred onto PVDF membrane (BioRad PteLtd). The membrane was probed separately using primary antibody specificfor acetylated H3, acetylated H4 or acetylated H2A (Upstate Pte Ltd).The detection antibody, goat anti rabbit antibody conjugated with HRPwas used according to the manufacturing instruction (Pierce Pte Ltd).After removing the detection antibody from the membrane, an enhancedchemiluminescent substrate for detection of HRP (Pierce Pte Ltd) wasadded onto the membrane. After removing the substrate, the membrane wasexposed to an X-ray film (Kodak) for 1 sec-20 mins. The X-ray film wasdeveloped using the X-ray film processor. The density of each bandobserved on the developed film could be analysed using UVP Bioimagingsoftware (UVP, Inc, Upland, Calif.). The values were then normalizedagainst the density of actin in the corresponding samples to obtain theexpression of the protein.

The results of immuno-blotting assay using histone deacetylase H3, H4and H2A antibodies are shown in Table 6.

TABLE 6 Histone acetylation activities Compound Histone-3 Histone-4Histone-2A 1 Active Active Active 2 Active Active Active 12 ActiveActive 17 Active Active Active 67 Active Active Active

These data demonstrate that compounds in this invention inhibit histonedeacetylases, thereby resulting in accumulation of acetylated histones.

Histone H3 Acetylation Assay—ELISA Approach

An Enzyme Linked Immunosorbent Assay (ELISA) can be applied to detectand quantify the acetylated histone3 (AcH3) in the protein lysateobtained from cancer cell lines treated with the HDAC inhibitors.

The ELISA assay was developed to detect the level of AcH3 from theColo205 colon cancer cell line treated with 10 μM HDAC inhibitorycompounds. The protein lysates from untreated and treated Colo205 wereobtained as previously described. The concentration of protein fromlysed cells was determined using the BCA method (Sigma-Aldrich Pte Ltd).

Different combinations of antibodies (see Table 7) that could be used asprimary antibody (capture antibody) or secondary antibody wereinvestigated in order to determine suitable antibodies as well as tooptimize antibody concentrations and assay conditions. It was found thatthe combination of mouse monoclonal antibody against H3 and rabbitpolyclonal antibody against AcH3 (Lys9/14) produced the best binding tothe antigens, either peptides or protein lysate from Colo205 coloncancer cell line treated with the HDAC inhibitors. No background wasobserved. The detection antibody used in this ELISA was donkey antirabbit conjugated with peroxidase.

To determine EC₅₀ where acetylated histone 3 was induced by 50%, Colo205cells was cultivated in 96 well plate at 1.5×10⁵ cells/well for 24 h.Colo205 cells were subsequently treated with HDAC inhibitors atdifferent doses (in duplicates, 9 doses treatment, 4-fold dilutions from100 μM). After treatment for 24 h, cells were lysed and the proteinconcentration was determined.

The ELISA plate (immulon 2HB plate, Biolaboratories Pte Ltd) was coatedwith 4 μg/ml of mouse monoclonal antibody against H3 at 4° C. overnight.After removed mouse monoclonal antibody against H3, the plate was washedwith PBS buffer containing 0.05% Tween-20 and blocked with thesuperblock solution (Pierce Pte Ltd) at 37° C., 1 h. The superblocksolution was removed and the plate was washed with the PBS buffercontaining 0.05% Tween. The AcH3 peptide, H3 peptide and the proteinlysates from treated Colo205 with the HDAC inhibitors were applied. Thecapture reaction between the primary antibody and the antigen, which ishistone 3 in the samples, was carried out at 37° C. for 1 h. Afterremoving the samples, the plate was washed with PBS buffer containing0.05% Tween. The secondary antibody, 0.5 μg/ml of rabbit polyclonalantibody against AcH3 (Lys9/14), was applied to detect the acetylationH3 in the samples at 37° C. for 1 h. After removing the secondaryantibodies, the plate was washed with PBS buffer containing 0.05% Tween.The detection antibody was applied to detect the secondary antibody thatcaptured AcH3 in the samples at 37° C. for 30 min. The substrate, 1-stepTurbo™B (Pierce Pte Ltd) was applied for 30 min until the color wasdeveloped. The reaction was stopped using 1M H₂SO₄. The absorbance wasmeasured at OD450 nm using Spectromax reader (Molecular DevicesCorporation, Sunnyvale, Calif.).

The standard curve was drawn and the concentration of AcH3 [(Lys9/14),μg/ml] in a sample was determined using the Softmax software inSpectromax. The amount of AcH3 in a sample was calculated based on thefollowing formula;

${{pg}\mspace{14mu} {of}\mspace{14mu} {AcH}\; 3{\left( {{Lys}\; {9/14}} \right)/{\mu g}}\mspace{14mu} {of}\mspace{14mu} {total}\mspace{14mu} {protein}\mspace{14mu} {total}} = \frac{\left( {{\mu g}\mspace{14mu} {of}\mspace{14mu} {AcH}\; 3\left( {{Lys}\; {9/14}} \right)\mspace{14mu} {in}\mspace{14mu} {the}\mspace{14mu} {assay}} \right)*10^{6}}{{\mu g}\mspace{14mu} {of}\mspace{14mu} {protein}\mspace{14mu} {in}\mspace{14mu} {the}\mspace{14mu} {assay}}$

Dose response curves were plotted to determine EC₅₀ values for thecompounds using XL-fit (ID Business Solution, Emeryville, Calif.).[Table 8]

TABLE 7 Antibodies used in the cross-species reactivity test and thecombination antibodies studies Detection antibody Antibodies used aseither primary or conjugated with HRP secondary antibody (horse radishperoxidase) Rabbit polyclonal antibody against AcH3 Donkey anti rabbit(Lys9/14; Upstate Pte Ltd), (Pierce Pte Ltd) Rabbit polyclonal antibodyagainst AcH3 Goat anti rabbit (Lys14; Upstate Pte Ltd), (Pierce Pte Ltd)Rabbit polyclonal antibody against AcH3 Goat anti mouse (Lys9, UpstatePte Ltd), (Pierce Pte Ltd) Goat polyclonal antibody against AcH3 Rabbitanti goat (Lys9/14, Santa Cruz Pte Ltd), (Pierce Pte Ltd) Goatpolyclonal antibody against H3 (N- Mouse anti goat 20, Santa Cruz PteLtd) (Pierce Pte Ltd) Mouse monoclonal antibody against H3 (Upstate PteLtd)

Data for selected compounds are presented in Table 8, as theconcentration effective for induction of acetylation of histone 3([AcH3(lys9/14)]) signal by 50% (EC₅₀).

TABLE 8 Compound Structure EC₅₀ (μM) 2

1.7 17

1.1 67

0.5

In Vivo Antineoplastic (or Anti-Tumor) Effect of HDAC Inhibiting Agents:

In data not shown, selected compounds were tested for maximal tolerateddose in normal mice and were found to be well tolerated by the mice withno obvious signs of toxicity or side effects in the dose range applied(which can be >200 mg/kg/day).

The efficacy of the compounds of the invention can then be determinedusing in vivo animal xenograft studies.

In these studies Female atymic nude mice (Harlan), 12-14 weeks of agewould be implanted subcutaneously in the flank with 5×10⁶ cells ofHCT116 or with 1×10⁶ cells of Colo205 human colon carcinoma suspended in50% Matrigel. When the tumor reaches the size 100 mm³, the xenograftnude mice would be paired-match into various treatment groups. Theselected HDAC inhibitors would be dissolved in appropriate vehicles,such as 10% DMA/10% Cremophore/80% water and administered to xenograftnude mice intraperitonelly by daily for 14 days. The dosing volume willbe 0.2-ml/20 g mouse. Paclitaxol, used as positive control, will beprepared for intravenous administration in 10% Ethanol/10%Cremophore/80% water. The dosing volume for Paclitaxol will be0.015-ml/g mouse. Tumor volume will be calculated every second day ofpost injection using the formula: Tumor volume (mm³)=(w²×l)/2, wherew=width and l=length in mm of an HCT116 or Colo205 carcinoma. Compoundsin this invention that are tested would show significant reduction intumor volume relative to controls treated with vehicle only. Theactivity of histone deacetylase when measured shall be reduced andresults in accumulation of acetylated histone relative to vehicletreated control group.

The details of specific embodiments described in this invention are notto be construed as limitations. Various equivalents and modificationsmay be made without departing from the essence and scope of thisinvention, and it is understood that such equivalent embodiments arepart of this invention.

What is claimed is:
 1. A method of treatment of a disorder selected fromthe group consisting of inflammatory disorders, immune system disorders,cardiovascular diseases, fibrotic diseases, vascular diseases, viraldiseases, neurological diseases, parasitic diseases and proliferativedisorders, the method comprising administering a therapeuticallyeffective amount of a compound of formula:

wherein R¹ is selected from the group consisting of: H, andunsubstituted or substituted C₁-C₁₀ alkyl, alkenyl, alkynyl, haloalkyl,heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl,cycloalkylalkyl, C₄-C₉ heterocycloalkylalkyl, arylalkyl, andheteroarylalkyl, each of which may be unsubstituted or substituted withone or more substituents independently selected from the groupconsisting of: halogen, ═O, ═S, —CN, —NO₂, alkyl, alkenyl, alkynyl,haloalkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl,hydroxy, hydroxyalkyl, alkoxy, alkoxyalkyl, alkylamino, acylamino,aminoalkyl, phenoxy, benzyloxy, alkylsulfonyl, arylsulfonyl,aminosulfonyl, —COOH, —C(O)OR⁵, —SH, and acyl; R² is selected from thegroup consisting of: H, halogen, and unsubstituted or substituted C₁-C₁₀alkyl, alkenyl, alkynyl, haloalkyl, heteroalkyl, cycloalkyl,heterocycloalkyl, aryl, heteroaryl, cycloalkylalkyl, C₄-C₉heterocycloalkylalkyl, arylalkyl, and heteroarylalkyl, each of which maybe unsubstituted or substituted with one or more substituentsindependently selected from the group consisting of: halogen, ═O, ═S,—CN, —NO₂, alkyl, alkenyl, alkynyl, haloalkyl, heteroalkyl, cycloalkyl,heterocycloalkyl, aryl, heteroaryl, hydroxy, hydroxyalkyl, alkoxy,alkoxyalkyl, phenoxy, benzyloxy, alkylamino, acylamino, aminoalkyl,alkylsulfonyl, arylsulfonyl, aminosulfonyl, —COOH, —C(O)OR⁵, —SH, andacyl; R³ is selected from the group consisting of H, C₁-C₆ alkyl, andacyl; X and Y are the same or different and are independently selectedfrom the group consisting of: H, halogen, —CN, —NO₂, —CF₃, C₁-C₄ alkyl,—COR⁵, —SR⁶, —OR⁶, and —NR⁷R⁸; R⁵ is C₁-C₄ alkyl; R⁶ is C₁-C₄ alkyl; R⁷and R⁸ are each independently selected from the group consisting of: H,C₁-C₄ alkyl, C₄-C₉ cycloalkyl, C₄-C₉ heterocycloalkyl, aryl, heteroaryl,arylalkyl and heteroarylalkyl; or a pharmaceutically acceptable saltthereof, to a patient in need thereof.
 2. A method according to claim 1wherein in the compound, R³=H.
 3. A method according to claim 1 whereinin the compound, X and Y=H.
 4. A method according to claim 1 wherein inthe compound R¹ is selected from the group consisting of: H, andunsubstituted or substituted hydroxyalkyl, C₁-C₁₀ alkyl, heteroalkyl,arylalkyl, heteroarylalkyl, and heterocycloalkyl, each of which may beunsubstituted or substituted as defined in claim
 1. 5. A methodaccording to claim 1 wherein in the compound R¹ is selected from thegroup consisting of: H, methyl; (pyridin-3-yl)methyl; 2-hydroxy-ethyl;2,3-di-hydroxy-propyl; and 3-hydroxy-propyl.
 6. A method according toclaim 1 wherein in the compound R² is selected from the group consistingof H, and unsubstituted or substituted C₁-C₁₀ alkyl, arylalkyl, aryl,heteroaryl, heteroalkyl, and cycloalkyl, each of which may beunsubstituted or substituted as defined in claim
 1. 7. A methodaccording to claim 1 wherein in the compound R² is selected from thegroup consisting of 2-phenyl-propyl, 2-phenyl-ethyl, 4-methoxy-phenyl,4-benzyloxy-3-methoxy-phenyl, pyridin-2-yl, and pyridin-4-yl.
 8. Amethod of claim 1 wherein the compound is selected from compounds, andtheir pharmaceutically acceptable salts, selected from the groupconsisting of


9. A method of claim 1 wherein the compound is selected from compounds,and their pharmaceutically acceptable salts, selected from the groupconsisting of


10. A method according to claim 1 wherein in the compound R¹ is selectedfrom the group consisting of: 2-(piperidin-1-yl)-ethyl;2-(pyrrolidin-1-yl)-ethyl; 2-diethylamino-ethyl; and3-dimethylamino-2,2-dimethyl-propyl.
 11. A method according to claim 1wherein the disorder is a inflammatory disorder or an autoimmunedisorder selected from the group consisting of Rheumatoid arthritis,Osteoarthritis, Juvenile chronic arthritis, Graft vs host disease,Asthma, Psoriasis, Ankylosing spondylitis, Crohn's diseases Inflammatorybowel disease, Lupus erythematosus and Chronic bronchitis.
 12. A methodaccording to claim 1 wherein the disorder is atherosclerosis.
 13. Amethod according to claim 1 wherein the disorder is a fibrotic disease.14. A method according to claim 14 wherein the fibrotic disease is liverfibrosis.
 15. A method according to claim 1 wherein the disorder is avascular disease.
 16. A method according to claim 15 wherein thevascular disease is selected from the group consisting of Human centralretinal vein occlusion, Van Hippel-Lindau disease and sporadivhemangioblastoma.
 17. A method according to claim 1 wherein the disorderis viral disease.
 18. A method according to claim 1 wherein the disorderis a neurological disease.
 19. A method according to claim 18 whereinthe neurological disease is selected from the group consisting ofHTLV-I-associated myelopathy and Alzheimer's Disease.
 20. A methodaccording to claim 1 wherein the disorder is a parasitic disorder.
 21. Amethod according to claim 1 wherein the disorder is a proliferativedisorder.
 22. A method according to claim 21 wherein the proliferativedisorder is cancer.
 23. A method according to claim 22 wherein thecancer is selected from the group consisting of colon cancer, ovariancancer, prostate cancer, breast cancer, lung cancer, liver cancer,pancreatic cancer, renal cancer, sarcoma, neuroblastoma, gastric cancer,multiple myeloma, myeloproliferative neoplasms and haematologic cancers.